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In some Asian countries, the seeds of Himalayan balsam or policeman's helmet, Impatiens glandulifera Royle are harvested as a food source. First introduced to Europe in 1838-1839, I. glandulifera quickly became a garden favourite and later on a prodigious weed. Nowadays I. glandulifera is highly invasive in almost whole Europe and occurs in various habitats. So far, little has been known about the genetic diversity of I. glandulifera in the Baltic region. The objective of this study was to evaluate the genetic variability of Lithuanian populations of I. glandulifera differing in geography or habitats by the randomly amplified polymorphic DNA (RAPD), using 8 primers. At the species level, all DNA bands (188) were polymorphic. Among populations of I. glandulifera genetic parameters ranged in the following intervals: 40-56% of polymorphic DNA bands, 0.115-0.165 for Nei's gene diversity, 0.179-0.255 for Shannon's information index. Pairwise genetic distances between populations ranged in the interval 0.088-0.259. AMOVA showed significant genetic differentiation of I. glandulifera populations in Lithuania (Φ PT = 0.511, p ≤ 0.01). Percentage of polymorphic DNA bands (generated by RAPD primers) of the populations correlated negatively with the site's mean temperature for vegetation season (r = -0.535, p < 0.015). Only for one population (Palanga population located near the Baltic sea) out of 20, significant relations were found between this population genetic and geographic distances to the other populations (r = -0.755, p < 0.001). Genetic distance-based cluster analyses for 400 individuals indicated 4 major groups of populations, among which there was no clear geographical pattern. O ur RAPD analyses indicate multiple introduction of this species in Lithuania. Presumably several different ways of invasion of I. glandulifera took place: natural run and predisposing it intentional and unintentional dispersal by human.
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ISSN 1392-3196   391
ISSN 1392-3196


RAPD analysis of genetic diversity among
Lithuanian populations of Impatiens glandulifera
1112
11
1


2

Abstract
   Impatiens glandulifera 
I. glandulifera
  I. glandulifera 
 I. glandulifera
           
I. glandulifera   
    I. glandulifera

           
I. glandulifera
p
rp

rp


       I. glandulifera       

 Balsaminaceae

Introduction
Impatiens-
         
 Impatiens walleriana I. hawkeri I. balsamina
I. glandulifera
    
I. glandulifera
  Impatiens
       
       
I. glandulifera    
      
 I. glandulifera     
      
 -


Impatiens

I. parvioraI. balfourii

       I. glandulifera
       
       

     
I. glanduliferaI. parviora
-


    I. glandulifera -
     
 
      
      
-

392
RAPD analysis of genetic diversity among Lithuanian populations of Impatiens glandulifera
     
 I. glandulifera
      




        -
      

  

-



    
       -

 







       
   Impatiens glandulifera  -

       

Materials and methods
Plant material.   I. glandulifera
        
      -
     
      

-


      

-
      
-
   Impatiens glanduli-
fera
     
 
-
-

     
  
    -



-



Table 1.Impatiens glandulifera
 
N














       631
       
      962 
       
       
   63    696
       696
       696
       696
       696
       696
       696
       696
       696
       
       696
       
      963 
      963 
      963 

       

      
          


DNA extraction and randomly amplied polymor-
phic DNA (RAPD) – polymerase chain reaction (PCR).-

-



    

      
-
ISSN 1392-3196   393
   -
       


Taq
2
Taq
RAPD analyses.    -
®-
       -
-
   

  



-
-


Statistical analyses.   

       

       
          

        




 .
-
-
GenAlEx Excel
-

G -


  
-
      


PopGene
-

-


 GenAlEx v 6.41 
  
-

Statistica v. 7.0

Results
Habitat features of populations selected for
RAPD analyses.    
 

 
     
-




-

 




 


      


      


Characteristics of bands generated by RAPD.
      
-
        

      


   
 







-



-

-
  -






Molecular variance among populations and
within populations    
      


 
   -

     
   -
  

RAPD analysis of genetic diversity among Lithuanian populations of Impatiens glandulifera
      

-
Table 2.
Impatiens glandulifera

















  32  21  
    19  
222    21  
      
269      
      
    29  
  36    
 
p
Table 3.Impatiens glandulifera -
I
 





I
 
    
    
    
    
    
    
    
    
    
    
    
    
    
    
    
    
    
    
    
 92   
    
 
p


-
         

-
-


      -


  
     -

 

     
    

-


       
-




ISSN 1392-3196   
Table 4.
Impatiens glandu-
lifera
  SS   

 19    

     
 399   
Note.
p
      
        
  
   -
    


       -
  


Table 5.
Impatiens glandulifera





12369 11 12 13   16   19 
1                   
2                  
3                  
                  
                  
6                  
                  
                  
9                  
                   
11                   
12                   
13                   
                   
                   
16                   
                   
                   
19                   
                   
Note.                 
               
                    

Figure 1.
Impatiens glandulifera
       



-





    
   
      -
   
       




       

-
396


      


 


         


RAPD analysis of genetic diversity among Lithuanian populations of Impatiens glandulifera
Figure 2.
       Impatiens
glandulifera
       
       
-
 
     
-

Figure 3.
Impatiens
glandulifera

  I. glandu-
lifera
  

rp



rp
   
      
       -

  

Discussion

Impatiens glandulifera-
-
I. glan-
dulifera
      


  
 I. glandulifera
     
-

I. glandulifera
-
-


I. glandu-
lifera

   

   I. glandulifera  

 

-

-



     
-
        

        
I. glandulifera

  
-
  
  I. glandulifera     

     
  


 
 
        
 -

 Impatiens     
   
-

 
      -

I. glandulifera
 

     -
     
ISSN 1392-3196   
     
-

        
  
 

      
 
-
     
Heracleum

-


-
       


I. glandulifera

       -
       
 I. glandulifera 
I. glandulifera
-


-
I. glandulifera-


        
      
   
-
 
       -
I. glandulifera
  

Lupinus polyphyl-
lus      Bunias orientalis


-
Impatiens glandulifera
-



I. glandulifera
 

       




Conclusions
   Impatiens glandulifera -

-


      
 -

-

Acknowledgements

  
      
    
Impatiens glandulifera



References
-
  Vaccinium oxycoc-
cus

          -
  -
Hypericum perforatum
 
Impatiens glanduliferaImpa-
tiens Roylei

Balsaminaceae-


        


-
 
-

   -
   Alliaria peti-
olata Brassicaceae 

-
  Erigeron an-
nuus

-
-
  

-
 Impatiens glandulifera Balsaminaceae

   
       Im-
patiens noli-tangere     Eustroma
reticulatum-

Impatiens glandulifera
-

Im-
patiens glandulifera-



    

          
Heracleum



RAPD analysis of genetic diversity among Lithuanian populations of Impatiens glandulifera

Im-
patiensBalsaminaceae 


    -
Phleum

-
-


Impatiens lateristachys Balsaminace-
ae 
-
 

   

  -
         

  -



-
Bunias orientalis
      

GenAlEx v. 6Excel
       

      -
   

-
   Impatiens glandulifera
-

Impatiens bal-
fourii -


 Impatiens
   



-
-

Im-
patiens -


       
Lupinus
polyphyllus      


-
-


ISSN 1392-3196


Bitinės sprigės (Impatiens glandulifera Royle) Lietuvos
populiacijų genetinė įvairovė pagal atsitiktinai pagausintą
polimornę DNR
111211
1
2
Santrauka
 Impatiens glandulifera 
             
   

         




rp
    

rp  
           



Balsaminaceae

... Plant leaf DNA was isolated from 105 individuals (5 individuals in each of 21 populations) by DNA purification kit #KO512 (Thermo Fisher Scientific, Vilnius, Lithuania), as documented earlier [62]. ...
... Standard parameters of genetic diversity, principal coordinate analyses (PCoA) for populations and individuals, and hierarchical AMOVA were obtained using GenAlEx v. 6.5. The unweighted pair group method with arithmetic mean (UPGMA)-based dendrogram was prepared by PopGene v. 1.32, similarly to that done for the data obtained by the other dominant multi-locus markers [35,62]. ...
... Our findings are similar to a Polish study which demonstrated exceptionally low polymorphism (6%) at four AFLP loci (two times less than in our study) for two populations [58]. Examination of a similar number (20 versus 21) of Lithuanian I. glandulifera populations at RAPD and SSR loci [62,74] showed much higher polymorphism compared to I. parviflora. Genetically, less polymorphic populations of I. parviflora were/are more successful invaders in Lithuania than I. glandulifera [4]. ...
Article
Full-text available
Currently, there is an increasing focus on understanding the interactions between genetic features of the invader and environmental factors that ensure the success of invasion. The objective of our study was to evaluate the genetic diversity of Lithuanian populations of highly invasive small balsam (Impatiens parviflora) by amplified fragment length polymorphism (AFLP) markers and to relate molecular data to biotope features defined by employing neighboring species of herbaceous plants. Low polymorphism of I. parviflora populations was observed at AFLP loci. Hierarchical analysis of molecular variance did not reveal differentiation of populations depending on biotope, geography, or road types. Bayesian analyses of AFLP data demonstrated many genetic clusters. Our results suggest multiple introductions of I. parviflora into Lithuania. The polymorphism of AFLP loci of populations significantly correlated with the total coverage by herbaceous plants in the sites. Defined by principal component analysis, the variability of study sites was most related to the coverage of herbaceous plants and least related to the molecular features of I. parviflora populations. The sites with I. parviflora were classified into agricultural scrubland, riparian forest, and urban forest biotopes. Of them, urban forest was distinguished by the highest coverage of I. parviflora and the lowest Ellenberg indicatory values for light, soil acidity, and richness in nutrients.
... Plant leaf DNA was extracted using DNA purification kit #K0512 (Thermo Fisher Scientific, Vilnius, Lithuania), following the instructions of the producer with some modifications (Zybartaite et al., 2011). ...
... Of the six tested primers, four (2-3, 5-6) generated clear and reproducible fragments of DNA and were used for population studies. Eight RAPD markers 222,250,269,340,474,and 516) applied for populations of I. glandulifera (Zybartaite et al., 2011) later were employed for Impatiens parviflora (Kupcinskiene et al., 2013). ...
... Congeneric for I. parviflora species, Impatiens glandulifera is also an annual plant existing in Europe already for the second century, known as top of widespread and worrying alien of the continent. Surprisingly, to date, molecular analyses of I. parviflora populations lag far behind I. glandulifera analyses performed in various countries by various types and sets of DNA markers (Walker, 2001;Provan et al., 2007;Walker et al., 2009;Zybartaite et al., 2011;Hagenblad et al., 2015;Jocienė, 2015;Nagy and Korpelainen, 2015;Cafa et al., 2020;Kurose et al., 2020;Korpelainen and Elshibli, 2021). ...
Article
Full-text available
Data on alien species show that plant invasions are caused by a complex combination of characteristics of invasive species (invasiveness) and characteristics of invaded environment (invasibility). Impatiens parviflora is one of Europe’s top invasive species. The present study aimed to evaluate molecular diversity of populations of highly invasive in Lithuania I. parviflora by applying several DNA markers and relating genetic parameters to abiotic and biotic environment. For sampling, urban forests, riparian forests, and agrarian shrublands were selected. Three different DNA-based techniques, Simple Sequence Repeats (SSR), Inter Simple Sequence Repeat (ISSR) markers, and Random Amplified Polymorphic DNA (RAPD), were used for detecting genetic variation between 21 populations. All population individuals were monomorphic and homozygotic for four loci and heterozygotic for one locus by SSR analysis. Hierarchical analyses of molecular variance at ISSR and RAPD loci revealed significant differentiation of populations depending on geographic zones of the country. Bayesian Structure analyses of molecular data demonstrated existence of many genetic clusters and this finding is in support to multiple introduction of the species. The polymorphism extent at ISSR loci was positively correlated with the total coverage of herbaceous plant species. The coverage by I. parviflora was negatively correlated with the total number of herbaceous species and light in the sites. Our results indicate that Lithuanian sites with I. parviflora might be distinguished by high soil nutrient levels. According to the principal component analysis, the coverage by I. parviflora was a more important variable of populations compared to molecular data or parameters of abiotic environment. In conclusion, complexity of invasibility and invasiveness factors determine the variability of I. parviflora sites, including genetic traits, coverage of invasive species and conditions of environment that were significant and interrelated.
... Additionally, human-mediated long-distance dispersal plays an important role (Walker et al. 2009, Zybartaite et al. 2011). With increasing time since invasion of a river, I. glandulifera spreads laterally from the river bank to adjacent, mostly riparian wet habitats, as well as upstream along the tributaries (Malíková and Prach 2010). ...
Thesis
Biological invasions are an important aspect of anthropogenic global change and considered to be one of the major threats to biodiversity worldwide. Invasive species can change native species abundance, community structure, and ecosystem processes in invaded communities. However, due to context-dependencies it is often difficult to generalize the impact of invasive species. The outcome of an invasion is for example influenced by species traits, the invaded ecosystem, and invasion stage. Developmental stages and site-specific conditions can modify the performance of the invader and its interaction with other organisms. Another challenge is to disentangle cause and effect of an invasion. An alien species can cause changes in a previously intact ecosystem, thus be the “driver” of the changes. Alternatively, it can be a “passenger” which is facilitated by previous ecosystem changes. Context-dependencies as well as causality of invasions are important issues to understand and evaluate invasions, and to develop more targeted management plans. Thus, I investigated context-dependencies and causality of the impact of invasions using the model plant species Impatiens glandulifera. It heavily invaded several habitat types in Central Europe, but its impact on native plant communities is rated ambiguously, pointing towards context-dependencies. In an experimental study under controlled conditions I assessed the competitive and allelopathic effects of I. glandulifera on native co-occurring plant species. I found that seedlings and juvenile plants were negatively affected by a combination of allelopathy and competition. Native species differed in their susceptibility to I. glandulifera, and juveniles were more affected than seedlings. 2-metoxy-1,4-naphtoquinone (2-MNQ), the supposed main allelochemical, led to minor reductions in plant growth, suggesting that it may not be the only allelopathic substance of I. glandulifera. With two field studies I tested whether this species-specific response of native plants to allelopathy and competition of I. glandulifera leads to changed community patterns. I performed an observational vegetation survey within heterogeneous riparian meadows and alder forests in Germany. The vegetation was recorded in summer and spring because of seasonal species turnover and thus potentially different impact of I. glandulifera. The abundance of I. glandulifera as well as its impact on native vegetation depended on the environmental conditions at a particular patch. Plant species α-diversity was found to be not affected, but native plant cover was reduced specifically to species and season. To see whether the impact of I. glandulifera is causal and thus I. glandulifera a driver of ecosystem changes, I conducted a field experiment within the same study sites. Invaded and uninvaded plots were compared with plots from which I. glandulifera was removed and plots where I. glandulifera was planted. A negative impact of planting I. glandulifera and a concurrent positive effect of removal on native vegetation biomass and Urtica dioica performance indicated a causal but low effect of I. glandulifera. Species α-diversity was again not affected. I suggest that I. glandulifera is a “back-seat driver” of changes, which is facilitated by previous ecosystem changes but is also a driver of further changes. In summary, I found that the impact of I. glandulifera depended on the native target species, their developmental stage, on the habitat, and season. Dominant plant species, especially U. dioica were most affected, species α-diversity was not affected, species composition only under specific conditions. I conclude that the impact of I. glandulifera on native vegetation is only minor to moderate. However, from literature it is known that I. glandulifera has a negative impact on several additional ecosystem properties such as mycorrhiza, soil properties, and herbivores. The results of my studies are also relevant to develop management strategies for I. glandulifera. Generally, eradication measures are not feasible at large scales because of the wide distribution of I. glandulifera. They should primarily be applied to sites which are valuable in terms of nature conservation and to sites, where the strongest impact of I. glandulifera is to be expected. This is the case in habitats with high light conditions in combination with high soil moisture. Attention should also be paid to habitats with distinct spring communities, because they were also affected by I. glandulifera. The understanding that I. glandulifera is not a clear driver of ecosystem changes, but has some characteristics of a back-seat driver, indicates that removal is not sufficient for management of an I. glandulifera population. Additionally, habitat-restoration is required to counter those changes that benefited the invasion of I. glandulifera.
... Multiple introductions of I. glandulifera into Europe have been suggested, and seem supported by observed genetic clustering shown among countries (Hagenblad et al., 2015;Kupcinskiene et al., 2015;Nagy and Korpelainen, 2015), but in some instances also within countries (Hagenblad et al., 2015;Kurose et al., 2020;Love et al., 2013;Nagy and Korpelainen, 2015;Zybartaite et al., 2011). This diverse population origin seems furthermore to be derived from both India and Pakistan Nagy and Korpelainen, 2015). ...
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This paper presents all current knowledge on the biology of the invasive therophyte Impatiens glandulifera Royle (Himalayan Balsam), and covers aspects of taxonomy, morphology, distribution, habitat requirements, ecology, life cycle, genetics, history of invasive spread, ecological impact and management. Although a few review papers have been published on this species in previous decades, a great deal of insights have been gained in the last three decades, owing to the species’ notorious reputation as one of the most problematic invasive species in Europe. This study consequently focusses on this novel information, with a particular focus on information from Central Europe.
... This scenario is equivalent to the sequential introduction events with subsequent gene pool mixing that has been observed for several invasive alien species [6,10,14]. Also note that the occurrence of populations with high genetic diversity in different parts of I. glandulifera's invaded range, such as Finland [35] and Lithuania [37], might be partly caused by such gene flow and subsequent gene pool mixing events. Especially the mixing of the genetically very dissimilar Stockholm populations [7], with the more southern populations could result in strong increases in local population genetic diversity. ...
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Background: The observation that many alien species become invasive despite low genetic diversity has long been considered the ‘genetic paradox’ in invasion biology. This paradox is often resolved through the temporal buildup genetic diversity through multiple introduction events. These temporal dynamics in genetic diversity are especially important for annual invasive plants that lack a persistent seed bank, for which population persistence is strongly dependent on consecutive seed ‘re-establishment’ in each growing season. Theory predicts that the number of seeds during re-establishment, and the levels of among-population gene flow can strongly affect recolonization dynamics, resulting in either an erosion or build-up of population genetic diversity through time. This study focuses on temporal changes in the population genetic structure of the annual invasive plant Impatiens glandulifera across Europe. We resampled 13 populations in 6 regions along a 1600 km long latitudinal gradient from northern France to central Norway after 5 years, and assessed population genetic diversity with 9 microsatellite markers. Results: Our study suggests sufficiently high numbers of genetically diverse founders during population reestablishment, which prevent the erosion of local genetic diversity. We furthermore observe that I. glandulifera experiences significant among-population gene flow, gradually resulting in higher genetic diversity and lower overall genetic differentiation through time. Nonetheless, moderate founder effects concerning population genetic composition (allele frequencies) were evident, especially for smaller populations. Despite the initially low genetic diversity, this species seems to be successful at persisting across its invaded range, and will likely continue to build up higher genetic diversity at the local scale.
... Every single primer produced a unique band pattern which is different from other primers. By combining all three primers, a comprehensive and plausible results will be achieved (Zybartaite et al., 2011). Combining dataset can also be done among different markers, such as RAPD and microsatellite (Tripathi, Saini, Mehto, Kumar & Tiwari, 2012). ...
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Ciplukan (Physalis angulata) is a medicinal plant in Indonesia, belonging to the family of Solanaceae. Based on molecular phylogenetic analysis, this plant is relative to Ashwaganda (Withania somnifera), the famed South Asia medicinal plant touted to kill cancer cells. A study was conducted on genetic variation of Ciplukan using Random Amplified Polymorphic DNA (RAPD) marker. A total of 23 plants from Northern, Southern, Central, Eastern, and Western part of Bandung were examined The RAPD analyses were performed using three selected random primers (OPA1, OPB17, and OPB10). Clustering analysis was conducted based on Unweighted Pair-Group Method with Arithmetic Average (UPGMA) using MEGA 4. Dendrogram showed that the sample plants were not grouped by their geographic localities, suggesting that a genetic interaction occurs among plants from five different locations. This result was also supported by the high level of estimated gene flow (Nm= 1.0919). It is likely due to the nature of self-incompatibility in Ciplukan which requires cross-pollination, creating a higher exchange of genes and leads to homogenization of genetic composition. Overall, these results indicated no genetic differentiation, meaning that all individuals remain taxonomically under the same species.
... Now this species is in a phase of intensive spread, which started in Lithuania few decades ago. Expansion of E. annuus as other neophytes might be promoted by global warming (prolonged vegetation season, rise of minimal temperatures) and anthropogenic factors, such as changes in agriculture practice in the past few decades, increasing international trade, extension of urban area (SCHMITZ et al., 2010;ZYBARTAITE et al., 2011). E. annuus in Lithuania is generally found in roadsides, abandoned fields, ruderal places, and lawns, near lakes, rivers and as weed in towns of the southern and southerneastern regions. ...
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This study was conducted to assess the clonal structure and genetic diversity of alien herbaceous plant species Erigeron annuus. The global warming and changes in agriculture practice in the past few decades were favourable for the expansion of this species in Lithuania. We used RAPD and ISSR assays to assess genetic variation within and among 29 populations of E. annuus. A total of 278 molecular markers were revealed. Our study detected reduced level of genetic diversity of invasive populations of E. annuus. Significant differences in DNA polymorphism among populations of E. annuus were also found. Some populations of this species are composed of genetically identical plants, while others were polymorphic. Clonal diversity of study populations ranged from 0.083 to 0.4 for both DNA marker systems. The Simpsons diversity index values ranged from 0.0 to 0.636. The average number of genotypes per population established using both assays was about 1.7. Out of 328 E. annuus individuals only 16 showed unique RAPD and 14 unique ISSR banding patterns. The remaining plants were clones of different size. The most common genotype of E. annuus identified in our study was represented by predominate in nine populations.
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Invasive plants are introduced multicellular organisms of the kingdom Plantae, which produce their food by photosynthesis. An invasive plant has the ability to thrive and spread aggressively outside its native range. A naturally aggressive plant may be especially invasive when it is introduced to a new habitat. The basic literature emphasizes mainly the ecological and environmental effects of invasive plants. Impacts of these plants on the food production have never been studied in details. The direct and indirect or potential effects of occurrence of invasive plants on food production have been analysed on basis of published data according to eight selected criteria: food, fodder for animals, food and drink additives, indirect support for food production, weeds on arable lands, meadow weeds, allergenic plants in food and toxic plants. The principal components analysis of habitat preferences of invasive plants in the Nitra river basin showed that the majority of invasive plants growing along rivers is edible (Fallopia spp., Helianthus tuberosus, Impatiens glandulifera) and invasive plants preferring drier agricultural fields or grasslands are toxic and/or allergenic with low or zero level of edibility (Ambrosia artemisiifolia, Heracleum mantegazzianum). The plants living in drier conditions may produce more toxins to protect the sources (eg. water) in their tissues than plants near water flows where there is abundance of sources.
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Distribution of Impatiens glandulifera in Lithuania is analysed. This cultivated and recently naturalized species is more frequent in the southern and eastern parts of the territory. Notes on biology of I. Glandulifera are presented. Communiteis in which this species is naturalized are described.
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We investigated the genetic characteristics of morphologically different wild cranberries (Vaccinium oxycoccos) from Čepkeliai using random amplified polymorphic DNA (RAPD) analysis. Twenty random primers were tested on genomic DNA extracted from 13 wild cranberry sprout clones. Reproducible amplification patterns were obtained using nine primers, which gave from 5 to 20 bands. The RAPD data revealed a high genetic variability within a wild cranberry population.
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A method is presented by which the gene diversity (heterozygosity) of a subdivided population can be analyzed into its components, i.e., the gene diversities within and between subpopulations. This method is applicable to any population without regard to the number of alleles per locus, the pattern of evolutionary forces such as mutation, selection, and migration, and the reproductive method of the organism used. Measures of the absolute and relative magnitudes of gene differentiation among subpopulations are also proposed.
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Describes the distribution, history, habitat, communities, autecology, reproduction and associated organisms of Indian balsam. Impatiens glandulifera is an alien, but completely naturalized in Britain on the banks of many types of waterways, waste ground, thin woodlands and occasionally mires. -from Authors
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