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The involvement of Narcissus hispanicus Gouan in the origin of Narcissus bujei and of cultivated trumpet daffodils (Amaryllidaceae)

June 2010
Anales del Jardín Botánico de Madrid 67(1)

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Naturalis Biodiversity Center

To investigate the origin of cultivated trumpet daffodils the genome size (2C-value) of more than 100 old and new cultivars were measured. A large number of concolored yellow tetraploid cultivars with large coronas seem to be derived from a doubling of chromosomes of a hybrid of the two species N. hispanicus Gouan and N. pseudonarcissus L. This can also still be recognized by the presence of a black spot at the top of the anthers in about 15 % of the tetraploid cultivars. Assuming N. pseudonarcissus as one of the parents of the allotetraploid cultivars, species of trumpet daffodils of section Pseudonarcissi investigated earlier were compared. Doubling of the nuclear DNA content of 23.8 pg of N. pseudonarcissus falls short of the values found for the trumpet daffodils. Species of trumpet daffodils of section Pseudonarcissi with a complementing higher amount of nuclear DNA like N. hispanicus with 25.8 pg, N. poeticus L. with 26 pg and the hexaploid N. pseudonarcissus ssp. bicolor (L.) Baker with 67.7 pg (Zonneveld,2008) and other species were evaluated. N. hispanicus with 25.8 pg clearly differs from N. pseudonarcissus with 23.8 pg and is accepted here also as a species. The nuclear DNA content of N. bujei (Fern. Casas) Fern. Casas with 30 pg fits with the (ancient) hybrid origin for N. bujei between N. longispathus Pugsley (36 pg) and N. hispanicus Gouan (25.8 pg). The tetraploids with white tepals and yellow corona's can be obtained by crossing a diploid N. pseudonarcissus with the hexaploid and bicolored N. pseudonarcissus ssp. bicolor (L.) Baker.

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The involvement of Narcissus hispanicus Gouan

in the origin of Narcissus bujei and of cultivated

trumpet daffodils (Amaryllidaceae)

B.J.M. Zonneveld

NCB Naturalis, section National Herbarium of The Netherlands, Leiden University,

P.O. Box 9514, 2300RA Leiden, The Netherlands.

Zonneveld@NHN.Leidenuniv.nl

Abstract

Zonneveld, B.J.M. 2010. The involvement of Narcissus hispani-

cus Gouan in the origin of Narcissus bujei and of cultivated trum-

pet daffodils (Amaryllidaceae). Anales Jard. Bot. Madrid 67(1):

29-39.

To investigate the origin of cultivated trumpet daffodils the

genome size (2C-value) of more than 100 old and new cultivars

were measured. A large number of concolored yellow tetraploid

cultivars with large coronas seem to be derived from a doubling

of chromosomes of a hybrid of the two species N. hispanicus

Gouan and N. pseudonarcissus L. This can also still be recog-

nized by the presence of a black spot at the top of the anthers in

about 15 % of the tetraploid cultivars. Assuming N. pseudonar-

cissus as one of the parents of the allotetraploid cultivars, species

of trumpet daffodils of section Pseudonarcissi investigated earli-

er were compared. Doubling of the nuclear DNA content of 23.8

pg of N. pseudonarcissus falls short of the values found for the

trumpet daffodils. Species of trumpet daffodils of section

Pseudonarcissi with a complementing higher amount of nuclear

DNA like N. hispanicus with 25.8 pg, N. poeticus L. with 26 pg

and the hexaploid N. pseudonarcissus ssp. bicolor (L.) Baker with

67.7 pg (Zonneveld,2008) and other species were eval-

uated. N. hispanicus with 25.8 pg clearly differs from N.

pseudonarcissus with 23.8 pg and is accepted here also as a

species. The nuclear DNA content of N. bujei (Fern. Casas) Fern.

Casas with 30 pg fits with the (ancient) hybrid origin for N. bujei

between N. longispathus Pugsley (36 pg) and N. hispanicus

Gouan (25.8 pg). The tetraploids with white tepals and yellow

corona’s can be obtained by crossing a diploid N. pseudonarcis-

sus with the hexaploid and bicolored N. pseudonarcissus ssp. bi-

color (L.) Baker.

Keywords: Narcissus hispanicus, daffodil origin, origin of

N. bujei.

Resumen

Zonneveld, B.J.M. 2010. Participación de Narcissus hispanicus

Gouan en el origen de Narcissus bujei y de los narcisos trompe-

ta cultivados (Amaryllidaceae). Anales Jard. Bot. Madrid 67(1):

29-39.

Para investigar el origen de los narcisos trompeta cultivados se

midió el tamaño del genoma (valor 2C) de más de 100 cultivares

viejos y nuevos. Un gran número de cultivares tetraploides ama-

rillos de color uniforme y con grandes coronas parecían derivar-

se de una duplicación de cromosomas de un híbrido de las dos

especies N. hispanicus Gouan y N. pseudonarcissus L. Esto tam-

bién se observa por la presencia de una mancha negra en la par-

te superior de las anteras en alrededor del 15 % de los cultivares

tetraploides. Tomando a N. pseudonarcissus por uno de los pa-

rentales de los cultivares alotetraploides, se compararon espe-

cies de narcisos trompeta de la sección Pseudonarcissi que ya

hubiesen sido investigados anteriormente. La duplicación del

contenido de ADN nuclear de 23,8 pg de N. pseudonarcissus

queda por debajo de los valores hallados en los narcisos trom-

peta. Se evaluaron otras especies de narciso trompeta de la

sección Pseudonarcissi con una cantidad más alta de ADN nu-

clear complementaria, tales como N. hispanicus, con 25.8 pg,

N. poeticus L., con 26 pg y el hexaploide N. pseudonarcissus ssp.

bicolor (L.) Baker, con 67.7 pg (Zonneveld, 2008), entre otras.

N. hispanicus, con 25.8 pg, se diferencia claramente de N. pseu-

donarcissus, con 23.8 pg, y también se acepta aquí como espe-

cie. El contenido de ADN nuclear de N. bujei (Fern. Casas) Fern.

Casas, con 30 pg, se adecua al hibridismo (antiguo) para N. bu-

jei entre N. longispathus Pugsley (36 pg) y N. hispanicus Gouan

(25,8 pg). Los tetraploides con tépalos blancos y coronas amari-

llas pueden obtenerse con el cruzamiento entre N. pseudonar-

cissus diploide y el hexaploide y bicolor N. pseudonarcissus ssp.

bicolor (L.) Baker.

Palabras clave: Narcissus hispanicus, origen de los narcisos

trompeta, origen de N. bujei.

Anales del Jardín Botánico de Madrid

Vol. 67(1): 29-39

enero-junio 2010

ISSN: 0211-1322

doi: 10.3989/ajbm.2219

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Introduction

Daffodil culture started with the import of large

numbers of bulbs collected from the field in the 16th

century. Especially the large concolored yellow N. his-

panicus (Gouan, 1773) was imported and consequent-

ly likely became exterminated in southern France and

most of the Pyrenees and later also most of Spain.

From these in culture at first triploids originated at the

end of the 19th century (Wylie, 1952) with N. ‘Golden

Spur’ found in the Estate Bakkershage in Wassenaar,

The Netherlands (De Mol, 1923). At roughly the same

time the triploids N. ‘Emperor’ and ‘Empress’ origi-

B.J.M. Zonneveld

nated with Backhouse UK (Wylie, 1952). At the start

of the 20th century the first tetraploids became avail-

able (N. ‘King Alfred’ 1899, N. ‘van Waverens Giant’,

1900) and since that time the tetraploid trumpet daf-

fodils have taken over the market.

Previous work in Hosta Tratt. (Zonneveld & Van

Iren, 2001), Galanthus L. (Zonneveld & al., 2003a),

Agapanthus L’Hér. (Zonneveld & Duncan, 2003b),

Nerine Herb (Zonneveld & Duncan, 2006), Narcissus

L. (Zonneveld, 2008) and Tulipa L. (Zonneveld, 2009)

has shown that nuclear DNA content of species and

cultivars can contribute to the taxonomy of species

and the origin of hybrids. The N. pseudonarcissus L.

Anales del Jardín Botánico de Madrid 67(1): 29-39, enero-junio 2010. ISSN: 0211-1322. doi: 10.3989/ajbm. 2219

Coll.# Species DNA Average St. Dev. Chrom. # Origin

in pg

PB436 N. hispanicus Gouan 25,9 25,8 0,3 2x= 14* S. de Nieves, Spain

D031 N. hispanicus Gouan 26,6 Cuenca, Spain

BZ 3-'08-1 N. hispanicus Gouan 25,5 Los Quejigales, Spain

BZ 3-'08-2 N. hispanicus Gouan 25,2 Los Quejigales, Spain

BZ 3-'08-3 N. hispanicus Gouan 25,6 Los Quejigales, Spain

BZ 3-'08-4 N. hispanicus Gouan 25,7 Los Quejigales, Spain

BZ 3-'08-5 N. hispanicus Gouan 25,5 Los Quejigales, Spain

N. hispanicus Gouan 25,6 ex H. Meeuwissen

BZ 3-'08-6 N. hispanicus Gouan 39,3 39,3 0,7 2n= 3x= 21 1 km from Los Quejigales

L16 N. 'Hispanicus Maximus' 36,5 35,4 0,9 2n= 3x= 21 H. Bulborum, Limmen

B35 N. 'Hispanicus Maximus' 36,1 ex C. Breed

N. 'Hispanicus Maximus' 34,7 ex H.Meeuwissen

N. 'Hispanicus Maximus' 35,3 ex C. Breed

N. 'Hispanicus Maximus' 34,4 ex B. Duncan ex JWB

A8715 N. bujei Fern. Casas 29,3 30,2 0,5 2n= 2x= 14 Cabra, Spain

D678 N. bujei Fern. Casas 30,5 Albacete, Spain

D701 N. bujei Fern. Casas 30,0 F. de la Pieta, Spain

D008 N. bujei Fern. Casas 30,1 ex M. Salmon SF137

BZ 3-2008 N. bujei Fern. Casas 29,5 10 km w v Cazorla

kw9613 N. bujei Fern. Casas 31,5 N. S del Sierra, Spain

N. pseudonarcissus L. 23,8 0,4 2x= 14* Zonneveld (2008)

N. abscissus (Haw.) Schult. f. 26,4 0,8 2x= 14& Zonneveld (2008)

N. moleroi Fern. Casas 26,1 0,4 2x= 14& Zonneveld (2008)

N. poeticus L. 26,0 0,5 2x= 14$ Zonneveld (2008)

N. cyclamineus DC 26,2 0,5 2x= 14+ Zonneveld (2008)

N. longispathus Pugsley 36,0 0,7 2x= 14* Zonneveld (2008)

N. nevadensis Pugsley 38,2 1,1 2x= 14* Zonneveld (2008)

N. pseudonarcissus ssp. bicolor 67,7 2,0 2n= 6x= 42 Zonneveld (2008)

Table 1. Narcissus species with their amount of nuclear DNA, standard deviation, chromosome number and origin. Chromosome

counts are from literature or derived from the genome size.

* Sañudo (1984); + Sañudo (1985); & Monserrat Martí & Vives (1991); $ Mehra & Sachdeva (1976).

2219 narcissus:Maquetación 1 18/06/2010 16:47 Página 30

complex has given rise in nature to many forms now

accommodated in eight subspecies (Zonneveld, 2008).

The genome sizes here measured show that N. hispa -

nicus Gouan is also of importance in the development

of yellow garden daffodils. It is moreover a good

species that can still be found in Southern Spain.

Moreover N. bujei (Fern. Casas) Fern. Casas is likely

derived from a cross between N. hispanicus and N. lon -

gispathus Pugsley. The tetraploids with white tepals

and yellow corona’s can be obtained by crossing the

diploid N. pseudonarcissus with the hexaploid N. pseu -

donarcissus ssp. bicolor (L.) Baker. Lastly, the te -

traploid cultivars with pink to orange color in the coro-

na show the influence of N. poeticus L.

The taxonomy of all species of Narcissus (Amarylli-

daceae) has been investigated recently by flow cytom-

etry (Zonneveld, 2008). The somatic nuclear DNA

contents (2C) were shown to range from 14 to 38

picogram for the diploids. The total number of Nar-

cissus species was determined as 36, nine more than in

Flora Europaea and they were divided up in two sub-

genera and eleven sections. Section Pseudonarcissi DC

was much more heterogeneous in nuclear DNA con-

tent than expected. Sixty five accessions of N. pseu -

donarcissus possessed, with 23.8 pg, similar amounts

of DNA. However several species from this section

were clearly distinctive in nuclear DNA content. It

runs from the diploid N. primigenius Fern. Casas with

21.7 pg to the also diploid N. nevadensis Pugsley with

38.2 pg. Also N. cyclamineus DC, N. abscissus (Haw.)

Schult. f. and N. moleroi Fern. Casas are with about

26 pg clearly different from N. pseudonarcissus. For

the first time, in eleven accessions, hexaploidy was

found in N. pseudonarcissus ssp. bicolor. A new sec-

tion Nevadensis Zonn. with 30-39 pg of nuclear DNA

was split off from the section Pseudonarcissi with now

21-27 pg.

The aim of the present study was the desire to know

whether nuclear DNA value was of use in determin-

ing the origin of trumpet daffodils and of the contro-

versial Narcissus bujei (Fern. Casas) Fern. Casas. This

is of value for the taxonomy of Narcissus and also for

improving commercial varieties.Moreover, important

pharmacological substances like galanthamines are

found in daffodils (Codina, 2002) making it worth-

while to be able to discriminate between the species

and show the origin of the main groups of cultivars.

Materials and methods

Plant material

Plant material of the species was mainly obtained

from the collections of J. Blanchard (UK) and D.

Origin of Narcissus bujei and trumpet daffodils

Donnison-Morgan (UK). However, most cultivars

were obtained from The Netherlands (KAVB Lisse,

Hortus Bulborum Limmen, C.P. Breed, C. van der

Veek, H. Meeuwissen, W. Lemmers, and S. de Groot)

and a few from the UK (J.M. Grimshaw, M. Salmon

and I. Young), from France (L. de Jager), from Ger-

many (G. Knoche) and from the USA (N. Wilson and

H. Koopowitz). Care was taken to ensure correct

identification of all material, relying also on the expert

opinions of J. Blanchard, D. Donnison-Morgan and

S. de Groot. In most cases fresh leaves were used from

the wild or available in the different collections.

Determination of nuclear DNA content

For the isolation of nuclei, about 0.5 cm of a fresh

full-grown leaf was chopped together with a piece of

Agave americana L. ‘Aureomarginata’ as internal stan-

dard. The nuclear DNA content (2C-value) of A. ame -

ricana was measured as 15.9 picogram (pg) per nucle-

us with human leukocytes (2C = 7 pg, Tiersch & al.,

1989) as the standard. The chopping was done with a

new razor blade in a Petri dish in 0.25 ml nuclei-isola-

tion buffer, with 0.01 % RNAse added (Zonneveld &

Van Iren, 2001b). After adding 1.8 ml propidium io-

dide solution (50 mg/l in isolation buffer) the suspen-

sion with nuclei was filtered through a 30 µm nylon

filter. The fluorescence of the nuclei was measured, 30

and 60 min. after addition of propidium iodide, using

a PARTEC CA-II flow cytometer. The more DNA is

present in a nucleus, the higher is the intensity of the

fluorescence. The 2C DNA content of the sample was

calculated as the sample peak mean, divided by the

Agave peak mean, and multiplied with the amount of

DNA of the Agave standard. From chopping of a leaf

piece of 0.5 cm2about 50 000 nuclei could be isolated.

For each clone, two to six different runs (determina-

tions) with at least 3000-5000 nuclei were measured

with two runs from a single nuclear isolation.

Results and discussion

In Table 2 nuclear genome size is presented for

more than 60 trumpet daffodils. These were compared

with the genome sizes of all species (Zonneveld, 2008,

Table 1). It turns out that doubling of the nuclear

DNA content of 23.8 pg of N. pseudonarcissus falls

short of the values found for the trumpet daffodils

(Table 3). Morphological similar looking species of

trumpet daffodils of section Pseudonarcissi with a

com plementing higher amount of nuclear DNA like

N. hispanicus with 25.8 pg, N. poeticus with 26 pg and

the hexaploid N. pseudonarcissus ssp. bicolor with

67.7 pg (Zonneveld, 2008) were selected and data on

Anales del Jardín Botánico de Madrid 67(1): 29-39, enero-junio 2010. ISSN: 0211-1322. doi: 10.3989/ajbm. 2219

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B.J.M. Zonneveld32

Anales del Jardín Botánico de Madrid 67(1): 29-39, enero-junio 2010. ISSN: 0211-1322. doi: 10.3989/ajbm. 2219

Table 2. Narcissus cultivars with their amount of nuclear DNA, average, standard deviation, chromosome number, origin and remarks. Chromosome counts are from Mol, 1923; Nagao,

1933; Fernandes & Fernandes, 1946; Fernandes & Neves, 1971; Sañudo, 1985, and Brandham & Kirton, 1985.

Coll.# Colour White tepals + pink trumpet DNA Average St. dev Chrom. # Origin Parents

in pg

2lim1 2WP N. ‘Accent’ 49,7 49,4 1,1 2n = 4x = 28 H. Bulborum Limmen, NL

2lim2 2WP N. ‘Algarve’ 50,0 H. Bulborum Limmen, NL

2lim3 2WP N. ‘Arctic Char’ 49,3 H. Bulborum Limmen,NL

2lim4 2WP N. ‘Eastern Dawn’ 48,3 H. Bulborum Limmen, NL

2lim6 2WP N. ‘Rose Noble’ 48,8 H. Bulborum Limmen, NL

2lim7 2WP N. ‘Precocious’ 50,7 H. Bulborum Limmen, NL

2lim8 2WP N. ‘Truly Royal’ 49,8 H. Bulborum Limmen, NL

2lim9 2WP N. ‘Valinor’ 50,6 H. Bulborum Limmen, NL

2lim10 2WP N. ‘Violetta’ 49,4 H. Bulborum Limmen, NL

1WP N. ‘Chanson’ 47,3 ex C. Breed

White tepals + yellow trumpet H. Bulborum Limmen, NL

1WY N. ‘W.P. Milner’ 23,0 23,5 0,4 ex C. Breed

1WY N. ‘Princeps’ 23,4 ex H.Meeuwissen

1WY N. ‘Topolino’ 23,8 2x= 14 ex commerce

H3 1WY N. ‘W.P. Milner’ 23,8 2x= 14 ex H. Bot. Leiden

1WY N. ‘Empress’ 38,8 38,9 0,5 3x= 21+B H. Bulborum Limmen, NL pseudon. × bicolor 4x

2lim17 1WY N. ‘Bizerta’ 48,4 49,6 0,9 H. Bulborum Limmen, NL

2lim13 1WY N. ‘Drummerry’ 48,9 H. Bulborum Limmen, NL

2lim16 1WY N. ‘Monticello’ 49,0 H. Bulborum Limmen, NL

2lim14 1WY N. ‘Bar None’ 49,2 H. Bulborum Limmen, NL

2lim18 1WY N. ‘Irish Mist’ 49,4 H. Bulborum Limmen, NL

2lim19 1WY N. ‘Wahkeena’ 49,5 H. Bulborum Limmen, NL

2lim15 1WY N. ‘Cyros’ 49,7 H. Bulborum Limmen, NL

2 1WY N. ‘Glory of Noordwijk’ 49,9 H. Bulborum Limmen, NL

2lim12 1WY N. ‘Breton’ 49,9 H. Bulborum Limmen, NL

2lim20 1WY N. ‘Tudor Groove’ 50,1 H. Bulborum Limmen, NL

2lim11 1WY N. ‘Bravoure’ 51,9 H. Bulborum Limmen, NL

1WY N. pseudon. ssp. leonensis 64,0 64,0 1,2 Picos de Europa

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Origin of Narcissus bujei and trumpet daffodils 33

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Coll.# Colour Yellow tepals + yellow trumpet DNA Average St. dev Chrom. # Origin Parents

in pg

G15 1YY N. ‘Bowles Early Sulphur’ 22,6 23,8 0,1 2x= 14 ex commerce

1 1YY N. gayi (Henon) Pugsley 23,9 H. Bulborum Limmen,NL

7 1YY N. ‘Golden Spur’ 23,8 H. Bulborum Limmen,NL

5 1YY N. ‘Henry Irving’ 24,4 H. Bulborum Limmen,NL

3 1YY N. ‘Emperor’ 37,8 37,8 0,5 3x= 21 H. Bulborum Limmen,NL pseudon × bicolor? (Rix)

1YY N. ‘Golden Harvest’ 46,3 48,1 1,1 4x = 28 ex C. Breed G. Spur × K Alfred

2YY N. ‘Saint Keverne’ 46,5 ex C. Breed

1YY N. ‘Dutch master’ 46,9 ex C. Breed

1YY N. ‘King Alfred’ 48,9 ex C. Breed

1YYd N. ‘Golden Ducat’ 47,7 ex C. Breed

2YY N. ‘Carlton’ 47,6 ex C. Breed

1YY N. ‘Meldrum’ 48,5 ex C. Breed

2lim21 1YY N. ‘Golden Perfection’ 51,6 53,4 2,4 H. Bulborum Limmen,NL

1YY N. ‘Rijnveld’s Early Sensation’ 55,1 2n = 33 ex C. Breed

Other Cultivars

3WO N. ‘Aflame’ 48,8 ex C. Breed

4WY N. ‘Argent’ 24,4 ex C. Breed

J5 8YY N. ‘Avalanche of Gold’ 46,6 ex L de Jager Bill Welch × tazetta

7YY N. ‘Baby Boomer’ 31,2 KABV 10-4-08 Avalanche × jonq

7YY N. × intermedius = Baby Boomer? 31,2 KABV 10-4-08 Avalanche × jonq

7YY N. ‘Baby Moon’ 33,7 ex commerce jonq minor × jonq

7YY N. ‘Baby Moon’ 67,5 ex commerce jonq minor × jonq

3YO N. ‘Bath’s Flame’ 37,2 ex C. Breed

4WY N. ‘Bridal Crown’ 28,2 2x = 17 ex commerce sport of l’Innocence’

L22 1WW N. ‘Celilo’ 49,0 H. Bulborum Limmen,NL Petsamo’ × ‘Beersheba’

11YOs N. ‘Centannees’ 49,2 H. Meeuwissen

2YO N. ‘Croesus’ 36,9 ex C. Breed

H2 7YW N. ‘Dickcissel’ 41,3 3x = 21 ex Hortus Bot. Leiden Binkie (2n = 28) × jonquilla

Table 2. (Continuación).

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B.J.M. Zonneveld34

Anales del Jardín Botánico de Madrid 67(1): 29-39, enero-junio 2010. ISSN: 0211-1322. doi: 10.3989/ajbm. 2219

Coll.# Colour Others cultivars DNA Average St. dev Chrom. # Origin Parents

in pg

B24 1YYd N. ‘Eystettensis’ 23,8 double flowers pseudon(niet × triandrus)

6YY N. ‘Februari Gold’ 37,8 3x= 21 ex commerce Golden Spur × cycl.

11WYs N. ‘Frileuse’ 36,4 3x= 21 ex commerce split crown

8WO N. Geranium 27,4 ex C. Breed

L19 8WO N. ‘Glorious’ 29,9 2x= 17 H. Bulborum Limmen,NL poeticus × tazetta

2lim23 8WY N. ‘Gran Primo Citronieri’ 47,0 2n= 32 H. Bulborum Limmen,NL

5YY N. ‘Hawera’ 24,3 2x= 14 ex commerce jonquilla × triandrus

2WY N. ‘Ice Follies’ 48,8 4x= 28 ex commerce

4YYd N. ‘Inglescombe’ 24,9 ex C. Breed

4WOd N. ‘Insulinde’ 37,2 ex C. Breed

6WY N. ‘Jack Snipe’ 39,0 3x= 21 ex commerce x cyclamineus

6YO N. ‘Jetfire’ 37,4 3x= 21 ex commerce x cyclamineus

L18 N. ‘Jumage’ 72,4 H. Bulborum Limmen,NL x tazetta

2YO N. ‘Killigrew’ 48,9 ex C. Breed

2lim5 2WO N. ‘Kilworth’ 49,9 H. Bulborum Limmen,NL

7YY N. ‘Kokopelli’ 41,7 ex H.Meeuwissen Sundial × fern.

L20 7YO N. ‘Lanarth’ 41,0 3x= 21 H. Bulborum Limmen,NL x jonquilla

L09 8WY N. ‘Laurens Koster’ 28,3 2x= 17 H. Bulborum Limmen,NL poeticus × tazetta

B5 6YY N. ‘Minicycla’ 23,8 ex C. Breed cyclamineus × astur.

H5 8WY N. ‘Minnow’ 28,0 ex Hortus Bot. leiden x tazetta

L05 13yy N. ‘Mite’ 24,5 H. Bulborum Limmen,NL obvallaris × cyclam.

J4 N. ‘Monarch × Northland’ 49,6 ex L. de Jager

H13 1WW N. ‘Mount Hood’ 49,3 4x= 28 ex Hortus Bot. Leiden

4WO N. ‘Orange Phoenix’ 24,8 ex C. Breed

11YYs N. ‘Pampaluna’ 48,5 ex C. Breed y,y split

6YY N. ‘Peeping Tom’ 37,1 3x= 21 ex commerce cycl × pseudonarc. 4x

7YWYW

N. ‘Pipit’ 42,8 ex commerce Binkie × jonquilla

5YY N. ‘Queen of Spain’ 33,5 ex C. Breed

3WY N. ‘Queen of the North’ 36,1 ex C. Breed

4YYd N. ‘Rip van Winkel’ 22,8 ex H.Meeuwissen

2WO N. ‘Royal Orange’ 49,6 H. Meeuwissen

Table 2. (Continuación).

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Origin of Narcissus bujei and trumpet daffodils

these and other related species are here presented

(Table 1). Species of other sections can be excluded as

these have mainly short trumpets and/or result (with

exception of N. poeticus) in sterile offspring. For com-

parison the results for 61 other cultivars are also given.

The establishment of N. hispanicus as a species

In 2008 I found a Narcissus in fair numbers in the

Parc Natural Los Quejigales in the Sierra de las

Nieves amidst Abies pinsapo Boiss. forests that turned

out to have a 2C-value of 25.8 pg. Other accessions

from the same area have similar 2C-values (Table

1). An exception was a plant found 10 km west of

Los Quejigales that with 39.3 pg clearly is a triploid.

Meadows (1972) found N. hispanicus in a remote just

opened spot in the Pyrenees, Val Cardos, Pleta de

Tornapigol, but this was not available for study. This

amount of on average 25.8 pg of nuclear DNA is clear-

ly different from the 23.8 pg for N. pseudonarcissus.

The plants from Los Quejigales conform with N. his-

panicus as described by Gouan (1773).It is character-

ized by big bright yellow colored flowers with spiraled

tepals, an ascending posture and a minute black spot

on the anthers. In 1788 Curtis described the same

species as N. major. Baker (1888) included it as a sub-

species of N. pseudonarcissus. The term ‘Major’ or

‘Maximus’ was later used for (m)any large yellow daf-

fodils. N. ‘Hispanicus Maximus’ in culture turns out

to be a triploid (Table 1). Narcissus hispanicus ‘Pineto-

rum’ PB436 with 25.8 pg was found by P. Bird in Sier-

ra de las Nieves and via M. Salmon grown by J. de

Groot from seed. It is presumably the same plant as

described by Smythies (1973) from the Sierra de las

Nieves in southern Spain as growing amidst Abies pin-

sapo. According to Smythies (1973) Willkomm col-

lected this species in 1861 in the Sierra de Yunquera

(now Sierra de las Nieves) at 1350 m and considered

them to be N. major. Webb (1980) considered N. his-

panicus as a form of N. pseudonarcissus ssp. major.

However Pugsley (1933), Fernandes (1968) and Fer-

nandes Casas (1986) regarded N. hispanicus as a good

species. Moreover, Graham & Barrett (2004) could

separate N. hispanicus from N. pseudonarcissus in

their phylogeny of Narcissus based on DNA se-

quences of two chloroplast genes. The arguments pre-

sented above supports species rank of N. hispanicus.

Its origin seems to be the Pyrenees, Southern France

(rare nowadays in both cases) and the Sierra de las

Nieves in southern Spain.

Species incorrectly thought to be N. hispanicus

Plants from Zezere Portugal described by Fernan-

des as N. hispanicus, N. confusus Pugsley from Sie-

Anales del Jardín Botánico de Madrid 67(1): 29-39, enero-junio 2010. ISSN: 0211-1322. doi: 10.3989/ajbm. 2219

Coll.# Colour Others cultivars DNA Average St. dev Chrom. # Origin Parents

in pg

BZ 3WY N. ‘Segovia’ 26,9 ex commerce rup watieri? × ?

2YY N. ‘Sir Watkin’ 36,3 ex C. Breed

N. ‘Snow Tube’ ? 48,2 KABV

H14 7YY N. ‘Sun Disc’ 25,9 2x= 14 ex Hortus Bot. Leiden rupicola × ( poeticus?)

7YY N. ‘Sundial’ 26,2 ex C. Breed poeticus × rupicola

2WY N. ‘Sweet Harmony’ 50,2 H. Meeuwissen

G16 YYd N. ‘Telemonius Plenus’ 24,3 2x= 14 ex J. Grimshaw = ’Van Sion’

12YY N. ‘Tete-a-Tete’ 39,7 3x= 24 ex commerce (CyclxSoleildÖr)nonredOP

3WO N. ‘The Star’ 25,2 ex C. Breed

3YO N. ‘Twinkle’ 36,4 ex C. Breed

1YYd N. ‘Wavertree’=’Giant’ 23,3 ex C. Breed coll. asturiensis

2WO N. ‘Will Scarlet’ 26,2 ex C van der Veek absc xpoet poetarum

YY N. × jonquilla 34,9 KABV

3WW N. ‘Xit’ 25,6 ex C. Breed

B34 4YYd N. ‘Yellow Cheerfulness’ 40,9 3x= 24 ex C. Breed Elvira’ ->’Y. Cheerfulness’

Table 2. (Continuación).

2219 narcissus:Maquetación 1 18/06/2010 16:47 Página 35

rra de Gredos, and from Embalse de Abraham, Sierra

Higuera, described by Fernández Casas as allied to

N. obvallaris (Henon) Pugsley are just plain N. pseu -

do narcissus with 23.8 pg (Zonneveld, 2008). Narcissus

obvallaris Salisb. and N. gayi Salisb., both of doubtful

origin, have an amount of nuclear DNA similar to

N. pseudonarcissus. However they have a black spot

on the anthers (Blanchard 1990; 1998). So it could be

suggested that occasionally N. pseudonarcissus has a

black spot. More likely, they show the influence of

N. hispanicus, although their amount of DNA does

not reflect that anymore.

The origin of N. bujei (Fern. Casas) Fern. Casas

Section Nevadensis Zonn. was separated from sec-

tion Pseudonarcissi based on its much higher amount

B.J.M. Zonneveld

of DNA of 36-38 pg (Zonneveld, 2008). At first, based

on 50% more DNA, they seemed triploids of N.

pseudonarcissus. However, they have as far as they

were counted 2n= 14 chromosomes (Sanudo, 1984)

and fully fertile pollen. So N. nevadensis Pugsley

(1933) with 38 pg and N. longispathus and its three

subspecies (Rois-Ruiz & al., 1998; Zonneveld, 2008)

with 36 pg are clearly separated from N. pseudonar-

cissus.

The N. bujei with 30.2 pg is described from the Ser-

rania de Ronda, southern Spain. It is clearly separate

in nuclear DNA content but hardly morphologically

from both N. pseudonarcissus and N. longispathus or

N. nevadensis. However, the latter two are character-

ized by having usually more than one flower to a stem.

Narcissus bujei was first described as a variety of

N. longispathus (N. longispathus var. bujei Fern. Casas:

“amicissimo Eugenio Domínguez Vilches, familia-

riter nobis ‘Buje’, ex animo dicata”, the orthography

therefore is with an “j”, not a “g”), later as a variety of

N. hispanicus and then as a species all by the same au-

thor (Fernández Casas, 1986, “bugei”), who in 2000

changed his mind again and made it a subspecies of N.

hispanicus. Both N. bujei and N. hispanicus are de-

scribed as having bright yellow flowers, with spirally

twisted tepals and leaves and a black spot on the

anthers. These data fit with the (ancient) hybrid origin

for N. bujei (30.2 pg) between N. longispathus (36 pg)

and N. hispanicus (25.8 pg).

The origin of cultivated trumpet daffodils

In the most recent taxonomical treatment of Nar-

cissus (Zonneveld, 2008) seven species are attributed

to the section Pseudonarcissi DC. with a 2C-value of

about 23.8 pg for all subspecies of N. pseudonarcissus.

The calculations in Table 3 show that plain tetraploids

from N. pseusonarcissus (23.8 pg) will have a 2C-value

of 47.6 pg. The nuclear DNA content of the large

trumpet daffodils have higher values (Table 2). This

suggests that a species of trumpet daffodil with a

higher amount of nuclear DNA is also involved in the

formation of the trumpet daffodils that are grown

nowadays. Three morphological similar species of

trumpet daffodils of section Pseudonarcissi have a

2C-value of about 26 pg: N. abscissus Schult. f., N. mo -

leroi Fern. Casas and N. hispanicus Gouan. N. abscis-

sus has a cut-off corona, a character rarely encoun-

tered in today’s daffodils, whereas the rare N. moleroi

has pale colored flowers. So both seem hardly in-

volved in today’s breeding. Although N. cyclamineus

DC. and N. poeticus L. from section Narcissus (Zonn-

eveld, 2008) have a similar genome size of about 26

pg, they differ too much in flower morphology to con-

Anales del Jardín Botánico de Madrid 67(1): 29-39, enero-junio 2010. ISSN: 0211-1322. doi: 10.3989/ajbm. 2219

Diploids Genome size (2C)

calculated, in pg

N. pseudonarcissus (measured) 23,8

N. hispanicus (measured) 25,8

N. pseudo × hisp 24,8

Triploids

N. pseudo × pseudo × pseudo 35,7

N. (pseudo × hisp ) × pseudo 36,7

N. (pseudo × hisp ) × hisp 37,7

N. hisp × hisp × hisp 38,7

Tetraploids

N. pseudo × pseudo × pseudo 47,6

× pseudo

N. pseudo × pseudo × pseudo × hisp 48,6

N. pseudo × hisp × pseudo × hisp 49,6

N. pseudo × hisp × hisp × hisp 50,6

N. hisp × hisp × hisp × hisp 51,6

Measured

N. hispanicus 'Maximus'.Hort. 36,3

N. 'Emperor' 37,8

N. 'Empress' 38,9

N. 'King Alfred' 49,0

N. Yellow, Yellow Corona (Table 2) 48,1

N. White, Yellow Corona (Table 2) 49,6

N. White, Pink Corona (Table 2) 49,4

Table 3. Calculated genome sizes of Narcissus species, hybrids

and cultivars compared with the actual measurements.

2219 narcissus:Maquetación 1 18/06/2010 16:47 Página 36

sider them as being involved with the origin of the yel-

low trumpet daffodils. This leaves us with N. hispani-

cus. N. hispanicus is characterized by big bright yellow

colored flowers with spiraled tepals, an ascending

posture and minute black spots on the anthers. Apart

from the spiraled tepals these characters can be found

back in today’s cultivars. A plain tetraploid N. hispan-

icus (25.8 pg) will have a 2C-value of 51.6 pg, higher

than any of the cultivated trumpet daffodils. This sug-

gests that both N. hispanicus and N. pseudonarcissus

are involved. The diploid plant from Sierra de las

Nieves has a 2C-value of about 26 pg. The following

can be envisioned: this mountain range is one of the

few remaining places were N. hispanicus survived. N.

hispanicus has been imported repeatedly in the old

days. This might be due to the fact that it is not very

well adapted to culture in N. Europe and it might re-

sent the summer wetness and winter frosts. Despite

that and also despite its spiraled tepals, N. hispanicus

has several characters worthwhile to introduce. First

it has big bright yellow colored flowers, whereas those

of N. pseudonarcissus are usually whitish to pale yel-

low. Further, contrary to N. pseudonarcissus that usu-

ally has drooping flowers, the flowers have an ascend-

ing posture. The main character contributed by N.

pseudonarcissus maybe its hardiness and non-spiraled

tepals. The hybrid seedlings with spiraled tepals

where said to have “mill wings” and vehemently se-

lected against (W. Lemmers, pers. comm.). Moreover

the hybrid might have shown hybrid vigour. The

diploid plant here described under N. hispanicus has

been crossed in culture with N. pseudonarcissus. The

hybrid might have been partly fertile and, as is often

the case with partly sterile hybrids, might have formed

non-reduced ovules. These have given rise to the

triploid N. ‘Hispanicus Maximus’. The plant in cultu -

re as N. “hispanicus” turns out to be this triploid. The

2C-value of 36.3 pg leads to envision a non-reduced

diploid gamete of N. pseudonarcissus × N. hispanicus

with a haploid gamete of N. pseudonarcissus or N. his-

panicus. Around 1860 three English breeders found

triploid daffodils. They were bought by Barr who

brought them in circulation in 1875 (Wylie, 1952).

The varieties N. ‘Emperor’ and N. ‘Empress’ were

also triploid. In 1890 tetraploids arose from them like

N. ‘King Alfred’, that is supposed to be a hybrid of

N. ‘Empress’ with N. hispanicus (Williams, 1929), and

in The Netherlands the triploid N. ‘Golden Spur’ was

followed by the tetraploid N. ‘van Waverens Giant’.

However, the plant in culture as N. ‘Golden Spur’

turned out to be diploid (Table 2). In these cases the

involvement of N. hispanicus was already suggested at

the time when they appeared on the market.

Origin of Narcissus bujei and trumpet daffodils

The cultivars mainly from Hortus Bulborum, Lim-

men, The Netherlands with 48.1 pg and with yellow

tepals and trumpets most conveniently can be ex-

plained as being allotetraploids of N. pseudonarcissus

and N. hispanicus. Autotetraploid N. pseudonarcissus

would have a 2C-value of 47.6 pg and autotetraploid

N. hispanicus would have a 2C-value of 51.6 pg.

Counts of Melissa Reading and Boyce Tankersley in

the USA and myself in the Hortus Bulborum of

Limmen (NL) have shown that up to 15 % of the

narcissi with large trumpets (division 1 and 2) have

black spots at the tip of the anthers. These black spots,

the bright yellow colors and upright posture shows

the influence of N. hispanicus in today’s daffodil

breeding.

Daffodils with white tepals

and with yellow trumpets

These have a 2C-value of 49.6 pg (Table 2) and have

likely been derived from crosses between hexaploid

N. pseudonarcissus ssp. bicolor from the Pyrenees and

Southern France and/or ssp. leonensis (Pugsley) Ferd.

Casas & Laínz from Northern Spain and a diploid

species, giving a tetraploid in the first generation.

Both wild hexaploid daffodils have white tepals and

large yellow trumpets.

Daffodils with pink or orange to red corona’s

All tetraploid cultivars around with orange to pink

corona’s are derived from N. poeticus and especially

from the tetraploid form N. poeticus ‘Actaea’. Tetra -

ploids with white tepals and pink trumpets with large

corona’s have on average a 2C-value of 49.4 pg. These

are clearly derived from N. poeticus with a 2C-value of

26 pg or its tetraploid cultivar ‘Actaea’ with 52 pg and

N. pseudonarcissus. Fernandes (1968a; 1968b) re-

marks on the similarity of the karyotypes of N. poeti-

cus of section Narcissus and section Pseudonarcissi. As

the hybrid between these two seems to be the only fer-

tile intersectional hybrid one may wonder whether or

not these two sections should be united. This is also

indicated by the results of Graham & Barrett (2006).

The red or pink color in today’s daffodil corona’s that

is exclusively derived from N. poeticus and the black

spot on the anthers are both unique characters.How-

ever in the first case there was and still is a very strong

selection for the red color. It is very unlikely anyone

has given any attention to the minute black spot on

the anthers. The black spots are absent from the in-

vestigated cultivars that show the red color of N. poet-

icus or the white tepals and yellow corona’s derived

from N. pseudonarcissus ssp. bicolor.

Anales del Jardín Botánico de Madrid 67(1): 29-39, enero-junio 2010. ISSN: 0211-1322. doi: 10.3989/ajbm. 2219

2219 narcissus:Maquetación 1 18/06/2010 16:47 Página 37

In Table 3 2C-values of different combinations of

species are calculated. If these are compared with the

actual data found (Tables 1, 3) and also taking the

flower colors into account, it is clear that N. hispanicus

played a role in the tetraploid yellow cultivars, N. poe -

ticus in the cultivars with red or pink corona’s and N.

pseudonarcissus ssp. bicolor in the tetraploids with

white tepals and yellow corona’s.

This is the first time that the difference in nuclear

DNA content is shown between N. hispanicus and

N. pseudonarcissus and is used to explain the origin

of N. bujei and the yellow tetraploid cultivars. Also

the DNA hexaploidy of N. pseudonarcissus ssp. bico-

lor and its use in obtaining tetraploid bicolored trum-

pet daffodils was not recorded earlier. These data al-

lowed giving a firm footing to the supposed involve-

ment of these species in the origin of the trumpet daf-

fodils.

Acknowledgements

I like to thank for the contribution of plant material by the

donors listed above. Special thanks go to John Blanchard and

Derrick Donnison-Morgan who both provided most of the plant

material for the species and C. Breed, C. van der Veek, H.

Meeuwissen, and J. Zonneveld (Hortus Bulborum Limmen, NL)

who provided most of the cultivars. Moreover based on their ex-

tensive knowledge on the subject they were always willing to dis-

cuss Narcissus matters with me. Dr. J.F. Veldkamp, NCB Na-

turalis, Leiden, The Netherlands, critically read a draft of the

manuscript.

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Received: 9-II-2009

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Full-text available

Feb 2020

The ploidy level can be useful in evaluating reproductive and somatic compatibility, an important parameter in scion and rootstock breeding programs. Evaluation of nuclear DNA content was performed in order to determine ploidy level of 31 rootstocks of genus Prunus for plum, sour cherry and sweet cherry. In the first stage of the research, the conditions of flow cytometry analysis with application of propidium iodide for DNA staining were optimized. Repeatable results and relatively good-quality histograms were obtained using Partec extraction buffer with 1% PVP (polyvinylpyrrolidone) addition, with incubation time from 50 to 90 min. The best internal standard for flow cytometry analysis proved to be tomato (Solanum lycopersicum L.) 2C = 1.96 pg for rootstocks with a nuclear DNA content from 0.55 to 1.64 pg, and soybean (Glycine max (L.) Merr.) 2.50 pg for rootstocks with a larger nuclear DNA contents from 1.8 to 2.3 pg. In five plum rootstocks, 2C values ranged from 0.61 to 0.67 pg indicating their diploid chromosome number. One rootstock was identified as tetraploid (2C = 1.34 pg), and four rootstocks as hexaploids owing to their DNA contents from 2.07 to 2.23 pg. One rootstock was considered a pentaploid due to its 2C value of 1.64 pg which was approximately two and a half times more than in diploid Prunus sp. In nine cherry rootstocks, 2C DNA values ranged from 0.74 to 0.86 pg indicating their diploid chromosome number. Nine cherry rootstocks were identified as triploids with their 2C DNA contents from 1.03 to 1.24 pg. Two cherry rootstocks were considered as tetraploids having 2C DNA of 1.39 pg. The ploidy level of cherry and plum rootstocks was evaluated in relation to morphological and agronomic traits. The tendency to increase the size of stomata and leaves along with an increase in the ploidy level was observed within Prunus rootstock genotypes; however, the correlations between these traits were not so evident. Therefore, stomata and leaf size cannot be considered as a morphological marker indicating ploidy level. The evaluated nuclear DNA contents / ploidy levels as well as stomata and leaf size are the additional descriptors of Prunus rootstocks that can be useful for identifying genotypes. Key words: flow cytometry, hexaploids, pentaploids, ploidy level, plum, cherry rootstocks, tetraploids, triploids.

Nuclear DNA Content and Ploidy Level of Apple Cultivars Including Polish ones in Relation to Some Morphological Traits

Article

Full-text available

Jan 2016
ACTA BIOL CRACOV BOT

Apple species and cultivars differ in nuclear (2C) DNA content and ploidy level. The majority of these genotypes are diploids, but there are some triploids and a few tetraploids. Nuclear DNA content is a specific feature and its flow cytometric evaluation can be helpful in differentiating taxa. For many apple genotypes – including all the Polish ones, these characteristics are not known. 2C DNA was evaluated in relation to leaf, flower, fruit, pollen grain and stomata sizes as well as to the flowering time for seventy genotypes (including 46 Polish cultivars) gathered in the gene bank of the Research Institute of Horticulture, Skierniewice, Poland. For standard cultivars with the known chromosome number, 2C value was 1.71 pg for diploid cultivar ‘Alwa’ (2n=2x=34), 2.55 pg for triploid ‘Boskoop’ (3x=51), and 3.37 pg for tetraploid genome (4x=68) of mixoploid ‘McIntosh 2x+4x’. In 61 cultivars (including 41 Polish ones), the nuclear DNA content ranged from 1.58 to 1.78 pg indicating their diploid chromo-some number. Five cultivars were identified as triploids (‘Bursztówka Polska’, ‘Pagacz’, ‘Rapa Zielona’, ‘Rarytas Śląski’ and ‘Witos’) owing to their nuclear DNA amount ranging between 2.42 and 2.58 pg. Leaf, flower, fruit, stomata and pollen grain sizes were on average significantly larger in triploids. Thus, in 3x plants the mean leaf surface was 49.1 cm

The Amaryllidaceae alkaloids: biosynthesis and methods for enzyme discovery

Article

Full-text available

Jun 2016
Phytochemistry Rev

Amaryllidaceae alkaloids are an example of the vast diversity of secondary metabolites with great therapeutic promise. The identification of novel compounds in this group with over 300 known structures continues to be an area of active study. The recent identification of norbelladine 4′-O-methyltransferase (N4OMT), an Amaryllidaceae alkaloid biosynthetic enzyme, and the assembly of transcriptomes for Narcissus sp. aff. pseudonarcissus and Lycoris aurea highlight the potential for discovery of Amaryllidaceae alkaloid biosynthetic genes with new technologies. Recent technical advances of interest include those in enzymology, next generation sequencing, genetic modification, nuclear magnetic resonance spectroscopy, and mass spectrometry.

Cloning and Characterization of a Norbelladine 4′-O-Methyltransferase Involved in the Biosynthesis of the Alzheimer’s Drug Galanthamine in Narcissus sp. aff. pseudonarcissus

Article

Full-text available

Jul 2014
PLOS ONE

Galanthamine is an Amaryllidaceae alkaloid used to treat the symptoms of Alzheimer's disease. This compound is primarily isolated from daffodil (Narcissus spp.), snowdrop (Galanthus spp.), and summer snowflake (Leucojum aestivum). Despite its importance as a medicine, no genes involved in the biosynthetic pathway of galanthamine have been identified. This absence of genetic information on biosynthetic pathways is a limiting factor in the development of synthetic biology platforms for many important botanical medicines. The paucity of information is largely due to the limitations of traditional methods for finding biochemical pathway enzymes and genes in non-model organisms. A new bioinformatic approach using several recent technological improvements was applied to search for genes in the proposed galanthamine biosynthetic pathway, first targeting methyltransferases due to strong signature amino acid sequences in the proteins. Using Illumina sequencing, a de novo transcriptome assembly was constructed for daffodil. BLAST was used to identify sequences that contain signatures for plant O-methyltransferases in this transcriptome. The program HAYSTACK was then used to identify methyltransferases that fit a model for galanthamine biosynthesis in leaf, bulb and inflorescence tissues. One candidate gene for the methylation of norbelladine to 4'-O-methylnorbelladine in the proposed galanthamine biosynthetic pathway was identified. This methyltransferase cDNA was expressed in E. coli and the protein purified by affinity chromatography. The resulting protein was found to be a norbelladine 4'-O-methyltransferase (NpN4OMT) of the proposed galanthamine biosynthetic pathway.

Analyzing Narcissus genome compositions based on rDNA loci on chromosomes and crossing-compatibility of 16 cultivars

Article

Jun 2020
SCI HORTIC-AMSTERDAM

Sixteen Narcissus cultivars were analyzed for their loci of 45S rDNA and 5S rDNA on mitotic metaphase chromosomes using fluorescence in situ hybridization (FISH), their male fertility by testing their pollen germination and female fertility based on their crossability. The results and analysis showed that thirteen cultivars, including ‘Queen's Day’, ‘Pink Charm’, ‘Stadium’, ‘Mount Hood’, ‘Eline’, ‘Accent’, ‘Dutch Master’, ‘Flower Parade’, ‘Replete’, ‘Las Vegas’, ‘Flower Surprise’, ‘Ice Follies’ and ‘Easter Born’, were autotetrapliod (2n = 4x = 28 = EEEE) and they were obtained from chromosome doubling, because they had similar FISH karyotypes and their male and female fertility match with autotetraploid; For similar reasons, Chinese Narcissus ‘Jinzhanyintai’ was autotriploid (2n = 3x = 30 = AAA) caused by unilateral sexual polyploidization, ‘Pink Parasol’ was identified as allotetraploid (2n = 4x = 30 = CCDD), and ‘Bridal Crown’ had one A genome and one B genome, meaning it was selected from interspecific hybridization (2n = 2x = 17 = A + B). It was concluded that FISH with rDNA probes was powerful tool to analyze Narcissus genomes. The results would be valuable to guide breeding in Narcissus.

A three-genome five-gene comprehensive phylogeny of the bulbous genus Narcissus (Amaryllidaceae) challenges current classifications and reveals multiple hybridization events

Article

Aug 2017

Besides being one of the most popular ornamental bulbs in western horticulture, the Mediterranean genus Narcissus has been the subject of numerous studies focusing on a wide scope of topics, including cytogenetics, hybridization and the evolution of polymorphic sexual systems. Phylogenetic hypotheses based on chloroplast data have provided a backbone for the genus but a detailed phylogenetic framework is still lacking. To fill this gap, we present a phylogenetic study of the genus using five markers from three genomes: ndhF and matK (chloroplast DNA), cob and atpA (mitochondrial DNA), and ITS (nuclear ribosomal DNA). In addition, we use chromosome counts from 89 populations representing 69 taxa. All analyses confirm that Narcissus is monophyletic with two main lineages largely corresponding to subg. Hermione and subg. Narcissus, but with incongruences between organellar and nuclear ribosomal phylogenies. At the infrageneric levels, our phylogenetic results challenge well-established taxonomic groups, such as sect. Jonquillae, sect. Bulbocodii and sect. Pseudonarcissi, each of which contains at least two distinct lineages that do not constitute monophyletic groups, and highlight the influence of allopolyploid species in the monophyly of sections within subg. Hermione. The type of the genus and its section is also nested within sect. Pseudonarcissi supporting new nomenclatural changes. Our results also confirm the intersubgeneric hybrid nature of several hybrids including allopolyploids (e.g., N. dubius, N. tortifolius, N. miniatus). Morphological and cytogenetic evidence independently support the hypothesis that some of the incongruence can be attributed to hybridization, such as the splits of sect. Bulbocodii and sect. Pseudonarcissi or the disparate phylogenetic placements of sect. Aurelia and sect. Ganymedes. Together, this indicates a significant role for reticulate evolution in shaping the diversity of this genus. A Bayesian divergence time analysis suggests that the major diversification events took place during the Neogene and provides younger estimates for the main nodes than previous studies, which fit paleoclimatic and paleotectonic reconstructions of the western Mediterranean during this period.

Cytological Observations on some W. Himalayan Monocots V. Araceae

Article

Full-text available

Jan 1976

The cytology of nine W. Himalayan species belonging to three genera Arisaema, Sauromatum and Typhonium has been studied. There is considerable variation in chromosome size in the genus Arisaema. Diploid taxon of A. concinnum (2n=28) has been found to occur in the Kumaon hills. Bowden had earlier reported this species to be tetraploid with 2n=56. A. curvatum was found to possess 2n=28. Malik had earlier observed 2n=26 in this species. Previously only the diploid form of A. jacquemontii was known to occur (Malik 1961). Present investigations have revealed the existence of tetraploids based on x=13 in the Simla hills. Sauromatum venosum has 2n=26 and possesses a symmetrical karyotype. The base number x=13 is suggested for the genus.

B.J.M. Zonneveld. 2008. The systematic value of nuclear DNA content for all species of Narcissus L. ( Amaryllidaceae). Plant Syst. Evol. 275 1,2: 109-132.

Article

Full-text available

Oct 2008

B.J.M. Zonneveld

The taxonomy of all species of Narcissus (Amaryllidaceae), an important horticultural crop, has not been investigated recently. As a new approach, genome size was determined by flow cytometry with propidium iodide from 375 accessions. The somatic nuclear DNA contents (2C) were shown to range from 14 to 38pg for the diploids. Narcissus assoanus and N. gaditanus are, based on their nuclear DNA content, removed from section Apodanthi and placed in a new section Juncifolii. The different ploidy levels and species involved were entangled for N . “fernandesii” s.l. and a new allotetraploid form is named here. Section Pseudonarcissus was much more heterogeneous in nuclear DNA content than expected. Sixty-five accessions of N. pseudonarcissus possessed, with 23.7pg, similar amounts of DNA. However, several species from this section were clearly distinctive in nuclear DNA content. It runs from the diploid N. primigenius with 21.7pg to the also diploid N. nevadensis with 38.2pg. Also N. abscissus and N. moleroi are with about 26pg clearly different from N. pseudonarcissus. For the first time, in 11 accessions, hexaploidy was found in N. pseudonarcissus ssp. bicolor. A new section Nevadensis with 30–39pg of nuclear DNA was split off from the section Pseudonarcissus with now 21–27pg. A nonoploid N. dubius with 96.3pg has by far the highest amount of nuclear DNA and can be calculated to have the highest ploidy ever reported in Narcisssus. The total number of Narcissus species was determined as 36, nine more than in Flora Europaea and they were divided up in two subgenera and 11 sections. Flow cytometry is shown to produce easily obtainable and original systematic data that lead to new insights. Genome size or C-value turns out to be one of the most salient features to define the status of the species in the genus Narcissus.

Taxonomic implications of genome size and pollen colour and vitality for species of Agapanthus L?H�ritier (Agapanthaceae)

Article

Full-text available

Oct 2003

Nuclear DNA content (2C) and pollen vitality and colour are used as new criteria to investigate all species of the genus Agapanthus LHritier. The species have the same chromosome number (2n=2x=30), with exception of four triploid plants found. The nuclear DNA content of the diploids, as measured by flow cytometry with propidium iodide, is demonstrated to range from 22.1–31.6 pg. This implies that the largest genome contains roughly 1010 more base pairs than the smallest. The species could be divided in two groups based on pollen colour and DNA content: a group with lilac pollen and a DNA content between 22.3 and 24.1 pg containing the species A. campanulatus Leighton, A. caulescens Sprenger and the rarer A. coddii Leighton, and a group with yellow/brownish pollen and a DNA content from 25.2–31.6 pg containing the species A. praecox Willd., A. inapertus Beauv. and A. africanus (L.) Hoffmanns. Four other taxa, recognized by Leighton (1965) are treated as follows: A. comptonii Leighton, has a nuclear DNA content similar to A. praecox and is considered to be a synonym of A. praecox subsp. minimus Leighton. A. walshii L. Bol., has with 31.6 pg the same high amount of DNA as A. africanus from the same area and is therefore renamed as a subspecies (A. africanus subsp. walshii (Leighton) Zonn. & Duncan comb. nov.). The nuclear DNA amounts of A. dyeri Leighton, including the geographically isolated plants from Mozambique, are shown to be identical to A. inapertus. Therefore A. dyeri is considered synonymous with A. inapertus subsp. intermedius Leighton. A. nutans Leighton is identical in DNA content to A. caulescens and is considered to be synonymous with that species. Concluding there are six species: A. campanulatus Leighton, A. caulescens Sprenger, A. coddii Leighton, A. praecox Willd., A. inapertus Beauv. and A. africanus (L.) Hoffmanns. Nuclear DNA content as measured by flow cytometry and pollen colour are shown to be relevant traits to throw light on the relationships between Agapanthus species.

A monograph of Narcissus, subgenus Ajax

Article

H.W. Pugsley

Contribution à la connaissance de la biosystématique de quelques espèces du genre Narcissus L

Article

Jan 1967

A. Fernandes

Cytological observations on some East-Himalayan Monocots

Article

Jan 1979

The Himalayas inhabit a fairly large number of representatives of the various monocotyledonous families, its eastern wing being comparatively much richer than the western (cf. Hooker 1894). Our previous communications in this journal provided cytological information on some W. Himalayan monocots (Mehra and Sachdeva 1975a,b; 1976a-e). The present paper deals with 21 taxa belonging to 18 species from the eastern part. Twenty one taxa belonging to 18 species of monocots are studied. Four species i.e. Alocasia acuminata (2n = 28), Cautleya lutea (n= 12, 13), Carex insignis (n=24) and C. longipes (n=21) are investigated for the first time. New chromosome numbers are reported for another three species, namely, Globba hookeri (n=12), Carex filicina (n=21) and Fimbristylis complanata (n=10). Heteromorphic chromosome pairs are noticed in Paris polyphylla (2n=20). © 1979, Japan Mendel Society, International Society of Cytology. All rights reserved.

The Chromosomes of Species, Hybrids and Cultivars of Narcissus L. (Amaryllidaceae)

Article

Jan 1987

Chromosome counts are reported for 46 taxa, 8 F1 hybrids and 230 cultivated varieties of Narcissus L. and the percentage pollen viability is given for most of them. B chromosomes were found in several species and cultivars. Chromosome numbers rise to the hexaploid level (2n = 42, 43) in species of the subgenus Narcissus, but the diploid (2n = 14) is the most commonly-occurring level of ploidy. In the subgenus Hermione diploid chromosome counts of 2n = 20 and 22 were obtained (x = 10, 11), in confirmation of reports by previous workers. In cultivars solely derived from the subgenus Narcissus eu- and aneu-tetraploids

(2n = 28 \pm 2)

greatly exceed diploids in frequency. Cultivars with complex ancestry involving x = 7 and x = 10 or 11 plants display a wide range of chromosome numbers, and the probable derivation of the chromosome complements of these cultivars and of other real and apparent aneuploids, together with those of some of the more interesting euploids, are given in the light of current knowledge. The levels of ploidy extant in Narcissus cultivars are compared and contrasted with those of a range of other genera in which many ornamental cultivars have also been produced.

The history of the garden Narcissi

Article

Aug 1952
HEREDITY

Ann P Wylie

An official journal of the Genetics Society, Heredity publishes high-quality articles describing original research and theoretical insights in all areas of genetics. Research papers are complimented by News & Commentary articles and reviews, keeping researchers and students abreast of hot topics in the field.

Genome Size and Pollen Viability as Taxonomic Criteria: Application to the Genus Hosta

Article

Feb 2008
PLANT BIOLOGY

Genome size (C values) and pollen viability staining were applied as new criteria to investigate the taxonomy of the genus Hosta Tratt. (Hostaceae). Nearly all species of the genus Hosta have the same basic chromosome number (2n = 2x = 60). However, the nuclear DNA contents, as measured by flow cytometry with propidium iodide, could be demonstrated to range between 17.2 to 26.6 pg. This implies that the largest genome contains roughly 1010 more base pairs than the smallest. Therefore, nuclear DNA content is a very relevant taxonomic trait that can be measured simply by flow cytometry. In addition, differences in overall DNA composition were demonstrated by comparing to DAPI fluorescence. In general, genome size data confirmed the division into three subgenera. The geographical distribution of genome sizes indicates the migration pattern of Hosta throughout East Asia. The species belonging to the mainly Korean subgenus Bryocles, with a low nuclear DNA content (17.2 - 19.3 pg), can now largely be distinguished from the mainly Japanese species of the subgenus Giboshi (21.3 - 26.5 pg). The exception is H. longissima, that with only 19.6 pg provides a nice example of a decrease in DNA content. On the mainland, as well as on Honshu, species with increased and decreased DNA content have evolved independently. The usefulness of pollen viability to detect hybrids in Hosta was demonstrated in a large series of artificial crosses between bona fide species. Consequently, pollen viability was measured in all available Hosta described as species. Several had low pollen viability and were concluded to be hybrids. Morphology and DNA content confirmed this in most cases. The resulting 23 species approximate the number of Hosta species that follows from the combined studies by Fujita (197618) on the Japanese species and Chung (1991 a11) on the Korean species.

Three new species of Narcissus L. subgenus Ajax Spach (Amaryllidaceae), restricted to the meadows and forests of south‐eastern Spain

Article

Oct 1999
BOT J LINN SOC

This paper provides descriptions of three new Narissus species—Narcissus segurensis sp. nov., Nalcaracensis sp. nov. and N. yepesii sp. nov.—that are endemic to the mountains of south-east Spain (Sierras de Alcaraz and Segura) and grow in meadows or deciduous oak forests. Characters, both vegetative and floral, used for determining resemblances and differences with other known taxa are discussed.