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ABSTRACT: Oryza sativa and Brassica napus-two important crops for food and oil, respectively-share high seed yield as a common breeding goal. As a model plant, O. sativa genomics have been intensively investigated and its agronomic traits have been advanced. In the present study, we used the available information on O. sativa to conduct comparative mapping between O. sativa and B. napus, with the aim of advancing research on seed-yield and yield-related traits in B. napus. Firstly, functional markers (from 55 differentially expressed genes between a hybrid and its parents) were used to detect B. napus genes that co-localized with yield-related traits in an F(2:3) population. Referring to publicly available sequences of 55 B. napus genes, 53 homologous O. sativa genes were subsequently detected by screening, and their chromosomal locations were determined using silico mapping. Comparative location of yield-related QTL between the two species showed that a total of 37 O. sativa and B. napus homologues were located in similar yield-related QTL between species. Our results indicate that homologous genes between O. sativa and B. napus may have consistent function and control similar traits, which may be helpful for agronomic gene characterization in B. napus based on what is known in O. sativa.
Journal of Genetics 08/2012; 91(2):163-70. · 1.09 Impact Factor
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ABSTRACT: Yellow seed color, which results from a thinner seed coat, is associated with improved feed quality of rapeseed (Brassica napus L.) meal and increased oil and protein content. As this trait follows various genetic models under different genetic backgrounds,
a study was performed in two genetic backgrounds to gain a better understanding of the genetic mechanisms underlying yellow
seed color. The quantitative trait locus (QTL) analysis was undertaken using two crosses, Quantum ×No. 2127-17 (HZ-1) and
No. 2127-17×94,570 (HZ-2). In the HZ-1 population, three putative QTL were detected in linkage groups N18, N5, and N3, respectively.
For all of them, yellow seed color arose from the No. 2127-17 alleles. Of these QTL, the one in linkage group N18 (Bnsc-18a) explained more than half of the phenotypic variation. In the HZ-2 population, three QTL were found in linkage groups N9,
N18, and N8, respectively. Of these QTL, that in linkage group N9 (Bnsc-9a) explained more than half of the phenotypic variation, whereas the QTL Bnsc-18a had a low seed color value and explained only 9.03–11.72% of the phenotypic variation. Bulked segregant analysis (BSA) of
the extremes of a BC1 population derived from the cross of No. 2127-17×94,570 (HZ-3) identified one major gene that was
identical with the QTL Bnsc-9a detected in the HZ-2 population. The QTL Bnsc-18a was common in the HZ-1 and HZ-2 populations, and the others were population-specific. These results suggested that different
black-seeded forms had different seed color genes.
Keywords
Brassica napus
–Seed color–QTL mapping
Molecular Breeding 04/2012; 28(3):335-342. · 2.85 Impact Factor
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ABSTRACT: Yellow seed is an important trait inBrassica napus. To know the genet ic basis of yellow seed color inBrassica napus, we carried out genetic studies by using conventional genetics analyses. The conventional genetics was studied in generations
(F1 F2 reciprocal F2, BC1, and F23) ofB. napus derived from crosses between a yellow-seeded (No. 2127-17) and nine different black-seeded parents. The results indicated
that seed color was mainly controlled by the maternal genotype but influenced by the interact ion between the maternal and
endosperm and/or embryonic genotypes. In the combinations which included black-seeded lines SW0780, 94560, 94545 and 1141B,
the yellow seed is partially dominant over black with two or three dominance epistasis ratio. A dominant yellow-seeded gene
Y which exhibits epistatic effects on the two independent dominant black-seeded genes B and C was ident ified in DH line No.
2127-17. These observations are in agreement with our previous reports. But in the rests, including the crosses with HS No.4,
HS No. 3, XY No. 15, 94570 and ZS No. 10, the black seed color was dominant over yellow seed color. The inheritance of this
trait in the segregating populations fits the model of a digenic dominance epistasis or triplicate dominance epistasis. A
new locus was identified and designated as D: the dominant gene D for black seed color inhibits the dominant gene Y. Therefore,
in combination with the Y, B and C, we found that the seed color was influenced by at least four genes. Identifying seed color
genes and defining their inheritance should further our understanding of yellow seed color trait and facilitate development
of new and better yellow-seeded cult ivars ofBrassics napus.
Key words
Brassica napus
-seed color-inheritance model-epistasis
Genes & genomics 04/2012; 31(6):413-419. · 0.44 Impact Factor
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Zhen Huang,
Lu Xiao,
Xiaoling Dun,
Shengqian Xia,
Bin Yi,
Jing Wen, Jinxiong Shen,
Chaozhi Ma,
Jinxing Tu,
Jinling Meng,
Tingdong Fu
[show abstract]
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ABSTRACT: Both the pollination control system and genetic distance are major factors in the utilization of crop heterosis. The recessive
genic male sterile line (RGMS) 7-7365A (Bnms3ms3ms4ms4) has been widely applied to hybrid seed production because it can generate a completely male sterile population by crossing
with the 7-7365C temporary line (Bnms3ms3rfrf). In this study, the sterile genes of 7-7365A were transferred to the new Brassica napus lines 7-749 and 7-750 with a high content of subgenomes by backcross breeding. We used the amplified fragment length polymorphism
(AFLP) technique combined with bulk segregant analysis (BSA) to identify markers linked to the BnMs4 gene. Twelve AFLP markers linked to the BnMs4 gene were identified. Of them, SA06MG09 and P08MG16 were the closest makers, which were on either side of the gene at a distance
of 0.9 and 0.8cM, respectively. Twenty AFLP primer combinations were used to screen the F2, BC1F3, and BC2F4 populations
from the breeding program, and the markers linked to the BnMs3 and BnMs4 genes were used to screen the BC2F4 populations. As a result, we obtained two types of improved sterile lines, 7-749A and
7-750A, and their indexes of subgenomic components (ISG) were 44.2–49.8 and 20.2–26.6%, respectively. The combining ability
analyses of seed yield character were conducted in the crosses from the three sterile lines and ten restorers within a random
block design in three environments for two successive years. The general combining ability (GCA) of the two improved sterile
lines were significantly higher than the GCA of 7-7365A in every environment tested. The two improved sterile lines had stability
in seed yield, and they will be used in the future for hybrid seed production.
Keywords
Brassica napus
–Subgenomic heterosis–Combining ability–Marker-assisted selection
Molecular Breeding 04/2012; 29(1):181-187. · 2.85 Impact Factor
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ABSTRACT: ‘SI1300’ is a self-incompatible Brassica napus line generated by introgressing an S haplotype from B. rapa ‘Xishuibai’ into a rapeseed cultivar ‘Huayou No. 1’. Five S-locus specific primer pairs were employed to develop cleaved amplified polymorphic sequences (CAPS) markers linked the S haplotype of ‘SI1300’. Two segregating populations (F2 and BC1) from the cross between ‘SI1300’ and self-compatible European spring cultivar ‘Defender’, were generated to verify the molecular
markers. CAPS analysis revealed no desirable polymorphism between self-incompatible and self-compatible plants. Twenty primer
pairs were designed based on the homology-based candidate gene method, and six dominant sequence characterized amplified region
(SCAR) markers linked with the S-locus were developed. Of the six markers, three were derived from the SRK and SP11 alleles of class II B. rapa
S haplotypes and linked with S haplotype of ‘SI1300’. The other three markers were designed from the SLG-A10 and co-segregated with S haplotype of ‘Defender’. We successfully combined two pairs of them and characterized two multiplex PCR markers which could
discriminate the homozygous and heterozygous genotypes. These markers were further validated in 24 F3 and 22 BC1F2 lines of ‘SI1300×Defender’ and another two segregating populations from the cross ‘SI1300×Yu No. 9’. Nucleotide sequences
of fragments linked with S-locus of ‘SI1300’ showed 99% identity to B. rapa class II S-60 haplotype, and fragments from ‘Defender’ were 97% and 94% identical to SLG and SRK of B. rapa class I S-47 haplotype, respectively. ‘SI1300’ was considered to carry two class II S haplotypes and the S haplotype on the A-genome derived from B. rapa ‘Xishuibai’ determines the SI phenotype, while ‘Defender’ carry a class I S haplotype derived from B. rapa and a class II S haplotype from B. oleracea. SCAR markers developed in this study will be helpful for improving SI lines and accelerating marker-assisted selection process
in rapeseed SI hybrid breeding program.
Molecular Breeding 04/2012; 21(3):305-315. · 2.85 Impact Factor
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ABSTRACT: Bleomycin, a glycopeptide antibiotic produced by the bacterium Streptomyces verticillus, has been demonstrated to be an effective mutagen in Arabidopsis thaliana. The objective of the present study was to determine the effect of bleomycin on embryo production and to assess the genetic
variation of the doubled haploid (DH) populations by amplified fragment length polymorphism (AFLP). The effects of bleomycin
on microspore embryogenesis and cell division were investigated using three concentrations of bleomycin in five semi-winter
genotypes of Brassica napus viz. T8, T10, B409, P30, and DH1142. Inclusion of bleomycin in the culture medium at a concentration of 0.1μgml−1 for 30min significantly improved embryo production and cell division in all five genotypes. Embryo production was induced
at rates two- and four-fold higher than controls after bleomycin treatment. Fifty plants regenerated by microspore embryogenesis
treated with bleomycin in addition to non-treated controls of T8, T10, and B409 were selected for AFLP analysis. The results
suggest that microspore culture is capable of producing 0.095–0.114% genetic variation, and there was no effect of bleomycin
treatment on genetic stabilisation of doubled haploid populations versus the non-treated control.
KeywordsAmplified fragment length polymorphism-
B. napus
-Doubled haploi-Glycopeptide antibiotic
Plant Cell Tissue and Organ Culture 04/2012; 101(1):23-29. · 3.09 Impact Factor
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ABSTRACT: Yellow-seeded Brassica napus was for the first time developed from interspecific crosses using yellow-seeded B. juncea (AABB), yellow-seeded B. oleracea (CC), and black-seeded artificial B. napus (AACC). Three different mating approaches were undertaken to eliminate B-genome chromosomes after trigenomic hexaploids (AABBCC) were generated. Hybrids (AABCC, ABCC) from crosses AABBCC × AACC, AABBCC × CC and ABCC × AACC were advanced by continuous selfing in approach 1, 2 and 3, respectively. To provide more insight into Brassica genome evolution and the cytological basis for B. napus resynthesis in each approach, B-genome chromosome pairing and segregation were intensively analyzed in AABCC and ABCC plants using genomic in situ hybridization methods. The frequencies at which B-genome chromosomes underwent autosyndesis and allosyndesis were generally higher in ABCC than in AABCC plants. The difference was statistically significant for allosyndesis but not autosyndesis. Abnormal distributions of B-genome chromosomes were encountered at anaphase I, including chromosome lagging and precocious sister centromere separation of univalents. These abnormalities were observed at a significantly higher frequency in AABCC than in ABCC plants, which resulted in more rapid B-genome chromosome elimination in the AABCC derivatives. Yellow or yellow-brown seeds were obtained in all approaches, although true-breeding yellow-seeded B. napus was developed only in approaches 2 and 3. The efficiency of the B. napus construction approaches was in the order 1 > 3 > 2 whereas this order was 3 > 2 > 1 with respect to the construction of yellow-seeded B. napus. The results are discussed in relation to Brassica genome evolution and the development and utilization of the yellow-seeded B. napus obtained here.
Theoretical and Applied Genetics 02/2012; 125(1):19-32. · 3.30 Impact Factor
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Shengqian Xia,
Ling Cheng,
Feng Zu,
Xiaoling Dun,
Zhengfu Zhou,
Bin Yi,
Jing Wen,
Chaozhi Ma, Jinxiong Shen,
Jinxing Tu,
Tingdong Fu
[show abstract]
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ABSTRACT: A recessive epistatic genic male sterile two-type line, 7365AB (Bnms3ms3ms4msRrfRf/BnMs3ms3ms4ms4RfRf), combined with the fertile interim-maintainer 7365C (Bnms3ms3ms4ms4rfrf) is an effective pollination control system in hybrid rapeseed production. We report an effective strategy used to fine map BnMs4 and BnRf. The two genes were both defined to a common microsyntenic region with Arabidopsis chromosome 3 using intron polymorphism (IP) markers developed according to Arabidopsis genome information and published genome organization of the A genome. The near-isogenic lines 7365AC (Bnms3ms3ms4ms4Rfrf/Bnms3ms3ms4ms4rfrf) of BnRf and 736512AB (Bnms3ms3Ms4ms4RfRf/Bnms3ms3ms4ms4RfRf) of BnMs4 were constructed to screen developed markers and create genetic linkage maps. Nine polymorphic IP markers (P1-P9) were identified. Of these, P2, P3, P4, and P6 were linked to both BnMs4 and BnRf with genetic distances <0.6 cM. Three simple sequence repeat markers, SR2, SR3, and SR5, were also identified by using public information. Subsequently, all markers linked to the two genes were used to compare the micro-collinearity of the regions flanking the two genes with Brassica rapa and Arabidopsis. The flanking regions showed rearrangements and inversion with fragments of different Arabidopsis chromosomes, but a high collinearity with B. rapa. This collinearity provided extremely valuable reference for map-based cloning in polyploid Brassica species. These IP markers could be exploited for comparative genomic studies within and between Brassica species, providing an economically feasible approach for molecular marker-assisted selection breeding, accelerating the process of gene cloning, and providing more direct evidence for the presence of multiple alleles between BnMs4 and BnRf.
Theoretical and Applied Genetics 01/2012; 124(7):1193-200. · 3.30 Impact Factor
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Xia Li,
Li Chen,
Meiyan Hong,
Yan Zhang,
Feng Zu,
Jing Wen,
Bin Yi,
Chaozhi Ma, Jinxiong Shen,
Jinxing Tu,
Tingdong Fu
[show abstract]
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ABSTRACT: Yellow seed is a desirable quality trait of the Brassica oilseed species. Previously, several seed coat color genes have been mapped in the Brassica species, but the molecular mechanism is still unknown. In the present investigation, map-based cloning method was used to identify a seed coat color gene, located on A9 in B. rapa. Blast analysis with the Arabidopsis genome showed that there were 22 Arabidopsis genes in this region including at4g09820 to at4g10620. Functional complementation test exhibited a phenotype reversion in the Arabidopsis thaliana tt8-1 mutant and yellow-seeded plant. These results suggested that the candidate gene was a homolog of TRANSPARENT TESTA8 (TT8) locus. BrTT8 regulated the accumulation of proanthocyanidins (PAs) in the seed coat. Sequence analysis of two alleles revealed a large insertion of a new class of transposable elements, Helitron in yellow sarson. In addition, no mRNA expression of BrTT8 was detected in the yellow-seeded line. It indicated that the natural transposon might have caused the loss in function of BrTT8. BrTT8 encodes a basic/helix-loop-helix (bHLH) protein that shares a high degree of similarity with other bHLH proteins in the Brassica. Further expression analysis also revealed that BrTT8 was involved in controlling the late biosynthetic genes (LBGs) of the flavonoid pathway. Our present findings provided with further studies could assist in understanding the molecular mechanism involved in seed coat color formation in Brassica species, which is an important oil yielding quality trait.
PLoS ONE 01/2012; 7(9):e44145. · 4.09 Impact Factor
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Zhengfu Zhou,
Xiaoling Dun,
Shengqian Xia,
Dianyi Shi,
Maomao Qin,
Bin Yi,
Jing Wen, Jinxiong Shen,
Chaozhi Ma,
Jinxing Tu,
Tingdong Fu
[show abstract]
[hide abstract]
ABSTRACT: 7365AB, a recessive genetic male sterility system, is controlled by BnMs3 in Brassica napus, which encodes a Tic40 protein required for tapetum development. However, the role of BnMs3 in rapeseed anther development is still largely unclear. In this research, cytological analysis revealed that anther development of a Bnms3 mutant has defects in the transition of the tapetum to the secretory type, callose degradation, and pollen-wall formation. A total of 76 down-regulated unigenes in the Bnms3 mutant, several of which are associated with tapetum development, callose degeneration, and pollen development, were isolated by suppression subtractive hybridization combined with a macroarray analysis. Reverse genetics was applied by means of Arabidopsis insertional mutant lines to characterize the function of these unigenes and revealed that MSR02 is only required for transport of sporopollenin precursors through the plasma membrane of the tapetum. The real-time PCR data have further verified that BnMs3 plays a primary role in tapetal differentiation by affecting the expression of a few key transcription factors, participates in tapetal degradation by modulating the expression of cysteine protease genes, and influences microspore separation by manipulating the expression of BnA6 and BnMSR66 related to callose degradation and of BnQRT1 and BnQRT3 required for the primary cell-wall degradation of the pollen mother cell. Moreover, BnMs3 takes part in pollen-wall formation by affecting the expression of a series of genes involved in biosynthesis and transport of sporopollenin precursors. All of the above results suggest that BnMs3 participates in tapetum development, microspore release, and pollen-wall formation in B. napus.
Journal of Experimental Botany 12/2011; 63(5):2041-58. · 5.36 Impact Factor
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Fupeng Li,
Chaozhi Ma,
Xia Wang,
Changbin Gao,
Jianfeng Zhang,
Yuanyuan Wang,
Na Cong,
Xinghua Li,
Jing Wen,
Bin Yi, Jinxiong Shen,
Jinxing Tu,
Tingdong Fu
[show abstract]
[hide abstract]
ABSTRACT: Sucrose is the primary photosynthesis product and the principal translocating form within higher plants. Sucrose transporters (SUC/SUT) play a critical role in phloem loading and unloading. Photoassimilate transport is a major limiting factor for seed yield. Our previous research demonstrated that SUT co-localizes with yield-related quantitative trait loci. This paper reports the isolation of BnA7.SUT1 alleles and their promoters and their association with yield-related traits.
Two novel BnA7.SUT1 genes were isolated from B. napus lines 'Eagle' and 'S-1300' and designated as BnA7.SUT1.a and BnA7.SUT1.b, respectively. The BnA7.SUT1 protein exhibited typical SUT features and showed high amino acid homology with related species. Promoters of BnA7.SUT1.a and BnA7.SUT1.b were also isolated and classified as pBnA7.SUT1.a and pBnA7.SUT1.b, respectively. Four dominant sequence-characterized amplified region markers were developed to distinguish BnA7.SUT1.a and BnA7.SUT1.b. The two genes were estimated as alleles with two segregating populations (F2 and BC1) obtained by crossing '3715'×'3769'. BnA7.SUT1 was mapped to the A7 linkage group of the TN doubled haploid population. In silico analysis of 55 segmental BnA7.SUT1 alleles resulted three BnA7.SUT1 clusters: pBnA7.SUT1.a- BnA7.SUT1.a (type I), pBnA7.SUT1.b- BnA7.SUT1.a (type II), and pBnA7.SUT1.b- BnA7.SUT1.b (type III). Association analysis with a diverse panel of 55 rapeseed lines identified single nucleotide polymorphisms (SNPs) in promoter and coding domain sequences of BnA7.SUT1 that were significantly associated with one of three yield-related traits: number of effective first branches (EFB), siliques per plant (SP), and seed weight (n = 1000) (TSW) across all four environments examined. SNPs at other BnA7.SUT1 sites were also significantly associated with at least one of six yield-related traits: EFB, SP, number of seeds per silique, seed yield per plant, block yield, and TSW. Expression levels varied over various tissue/organs at the seed-filling stage, and BnA7.SUT1 expression positively correlated with EFB and TSW.
Sequence, mapping, association, and expression analyses collectively showed significant diversity between the two BnA7.SUT1 alleles, which control some of the phenotypic variation for branch number and seed weight in B. napus consistent with expression levels. The associations between allelic variation and yield-related traits may facilitate selection of better genotypes in breeding.
BMC Plant Biology 11/2011; 11:168. · 3.45 Impact Factor
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ABSTRACT: Cytoplasmic male sterility (CMS) is a widespread phenomenon in higher plants, and several studies have established that this maternally inherited defect is often associated with a mitochondrial mutant. Approximately 10 chimeric genes have been identified as being associated with corresponding CMS systems in the family Brassicaceae, but there is little direct evidence that these genes cause male sterility. In this study, a novel chimeric gene (named orf288) was found to be located downstream of the atp6 gene and co-transcribed with this gene in the hau CMS sterile line. Western blotting analysis showed that this predicted open reading frame (ORF) was translated in the mitochondria of male-sterile plants. Furthermore, the growth of Escherichia coli was significantly repressed in the presence of ORF288, which indicated that this protein is toxic to the E. coli host cells. To confirm further the function of orf288 in male sterility, the gene was fused to a mitochondrial-targeting pre-sequence under the control of the Arabidopsis APETALA3 promoter and introduced into Arabidopsis thaliana. Almost 80% of transgenic plants with orf288 failed to develop anthers. It was also found that the independent expression of orf288 caused male sterility in transgenic plants, even without the transit pre-sequence. Furthermore, transient expression of orf288 and green fluorescent protein (GFP) as a fused protein in A. thaliana protoplasts showed that ORF288 was able to anchor to mitochondria even without the external mitochondrial-targeting peptide. These observations provide important evidence that orf288 is responsible for the male sterility of hau CMS in Brassica juncea.
Journal of Experimental Botany 11/2011; 63(3):1285-95. · 5.36 Impact Factor
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ABSTRACT: To explore a new way to modify the UPPP and to assess the operative efficacy of modified-coblation assisted UPPP (M-CAUP) on patients with severe obstructive sleep apnea-hypopnea syndrome (OSAHS).
A retrospective analysis was made on surgical therapeutic effect of M-CAUP performed in our hospital. Inclusion criteria: 87 adult inpatients with severe OSAHS were involved in the study whose preoperative AHI were beyond 30/h and LSaO2 were under or equal to 85%. All those had the primary level obstruction within the retropalatal region as determined by PSG and airway pressure fluctuation monitoring (ApneaGraph 200, MRA-Medical Ltd, Gloucestershire UK) . The surgical approach of M-CAUP: (1) bilateral tonsillectomy by using Arthrocare EVac 70 T&A Wand. (2) dissection of velum palatinum space by using Arthrocare EVac 70 T&A Wand, ablation of fat tissue to removed it from the space, avoiding injure levator palatini muscle and tensor palatini muscle and preserving musculus uvula, maintaining the normal structure of oropharynx. (3) ablation of channel of soft palate by using Arthrocare EVac 55 T&A Wand to reduce the volume of soft palate moderately. (4) plasty: suturing the tonsilar fossa to close the space, interrupted suture two sides of palatine arch and uvula. All patients were followed up for six to eighteen months postoperatively and received PSG.
After M-CAUP, the pharyngeal cavity was enlarged while the basic structure of oropharynx was maintained and no nasopharyngeal refluxing occurred. The subjective symptoms were also improved evidently after operation. After operation, the AHI and Epworth sleepiness scale (ESS) decreased while LSaO2 increased. The reduction of AHI and ESS had statistical significance (P < 0.01), and the rise of LSaO2 also had statistical significance (P < 0.01). The total effective ratio of the operation was 89.7%.
Despite of various treatment, the operative efficacy of conventional UPPP was almost not ideal on severe OSAHS due to complex pathogeny. It was proved that M-CAUP was an effective surgical treatment of severe OSAHS with less blood loss. The operation was minimally invasive and maintained the normal functions of palatopharynx in principle. It could be applied in clinical practice.
Lin chuang er bi yan hou ke za zhi = Journal of clinical otorhinolaryngology 10/2011; 25(20):929-32.
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ABSTRACT: Here, we describe the characteristics of a Brassica napus male sterile mutant 7365A with loss of the BnMs3 gene, which exhibits abnormal enlargement of the tapetal cells during meiosis. Later in development, the absence of the BnMs3 gene in the mutant results in a loss of the secretory function of the tapetum, as suggested by abortive callose dissolution and retarded tapetal degradation. The BnaC.Tic40 gene (equivalent to BnMs3) was isolated by a map-based cloning approach and was confirmed by genetic complementation. Sequence analyses suggested that BnaC.Tic40 originated from BolC.Tic40 on the Brassica oleracea linkage group C9, whereas its allele Bnms3 was derived from BraA.Tic40 on the Brassica rapa linkage group A10. The BnaC.Tic40 gene is highly expressed in the tapetum and encodes a putative plastid inner envelope membrane translocon, Tic40, which is localized into the chloroplast. Transmission electron microscopy (TEM) and lipid staining analyses suggested that BnaC.Tic40 is a key factor in controlling lipid accumulation in the tapetal plastids. These data indicate that BnaC.Tic40 participates in specific protein translocation across the inner envelope membrane in the tapetal plastid, which is required for tapetal development and function.
The Plant Journal 07/2011; 68(3):532-45. · 6.16 Impact Factor
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Xinhua Zeng,
Lixia Zhu,
Yanli Chen,
Liping Qi,
Yuanyuan Pu,
Jing Wen,
Bin Yi, Jinxiong Shen,
Chaozhi Ma,
Jinxing Tu,
Tingdong Fu
[show abstract]
[hide abstract]
ABSTRACT: In the present study, we have obtained one dwarf mutant (bnaC.dwf) from the Brassica napus inbred line T6 through chemical mutagen ethyl methanesulfonate (EMS). We have determined the phenotypic effects and genetic characteristics of dwarf mutant (bnaC.dwf). The dwarf mutant was insensitive to exogenous GA(3) for plant height, suggesting that it is significantly playing a crucial role in the gibberellins response pathway. Genetic analysis revealed that one recessive gene is responsible for controlling the phenotypic expression of dwarf mutant. Amplified Fragment Length Polymorphism (AFLP) technique was applied for selecting markers linked to the BnaC.DWF gene which assisted in screening of dwarf and normal individuals in the BC(4) population. We have screened 1,024 primer combinations and then identified nine AFLP markers linked to the BnaC.DWF gene. Identification and linkage of the markers were carried out by analysing 2,000 individuals from a larger population of the BC(4). Two markers EA10MC09 and EA12MC02 were located on the flanking region of the BnaC.DWF gene at a distance of 0.2 and 0.05 cM, respectively. Four AFLP markers EA09MG05, EA02MC07, EA01MC01 and EC04MC07 were successfully converted into Sequence Characterised Amplified Region markers namely SCA9G5, SCA2C7, SCA1C1 and SCC4C7. We further integrated BnaC.DWF linked Simple Sequence Repeat markers into two populations (Piquemal et al. Theor Appl Genet 111:1514-1523, 2005; Cheng et al. Theor Appl Genet 118:1121-1131, 2009). BnaC.DWF was mapped to the linkage region N18. The molecular markers developed from these investigations will greatly accelerate the selection process for developing dwarf varieties in B. napus by Marker Assisted Selection and genetic engineering.
Theoretical and Applied Genetics 09/2010; 122(2):421-8. · 3.30 Impact Factor
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Bin Yi,
Fangqin Zeng,
Shaolin Lei,
Yunin Chen,
Xueqin Yao,
Yun Zhu,
Jing Wen, Jinxiong Shen,
Chaozhi Ma,
Jinxing Tu,
Tingdong Fu
[show abstract]
[hide abstract]
ABSTRACT: S45A, a double recessive mutant at both the BnMs1 and BnMs2 loci in Brassica napus, produces no pollen in mature anthers and no seeds by self-fertilization. The BnMs1 and BnMs2 genes, which have redundant functions in the control of male fertility, are positioned on linkage groups N7 and N16, respectively, and are located at the same locus on Arabidopsis chromosome 1 based on collinearity between Arabidopsis and Brassica. Complementation tests indicated that one candidate gene, BnCYP704B1, a member of the cytochrome P450 family, can rescue male sterility. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) of the developing anther showed that pollen-wall formation in the mutant was severely compromised, with a lack of sporopollenin or exine. The phenotype was first evident at the tetrad stage (stage 7) of anther development, coinciding with the maximum BnCYP704B1 mRNA accumulation observed in tapetal cells at stages 7-8 (haploid stage). TEM also suggested that development of the tapetum was seriously defective due to the disturbed lipid metabolism in the S45A mutant. A TUNEL assay indicated that the pattern of programmed cell death in the tapetum of the S45A mutant was defective. Lipid analysis showed that the total fatty acid content was reduced in the S45A mutant, indicating that BnCYP704B1 is involved in lipid metabolism. These data suggest that BnCYP704B1 participates in a vital tapetum-specific metabolic pathway that is not only involved in exine formation but is also required for basic tapetal cell development and function.
The Plant Journal 09/2010; 63(6):925-38. · 6.16 Impact Factor
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ABSTRACT: Male sterility is an important contributor to heterosis in Brassica napus L. The B. napus line 7-7365ABC is a recessive epistatic genic male sterile (REGMS) three-line system. The 7-7365A line with the genotype Bnms3ms3ms4ms4RfRf is male-sterile, while the 7-7365B line with the genotype BnMs3ms3ms4ms4RfRf is male-fertile, and 7-7365C with homozygous recessive genotypes at the three loci shows male fertility because the loss function of Bnrf gene causes the inhibition of the genetic trait of the double mutant Bnms3 Bnms4. Histological studies addressing male sterility, transcriptional regulation pathways and the role of abscisic acid (ABA) in the anther development of REGMS plants are reported here. In the male-sterile line 7-7365A, tapetum cell and microspore mother cell separation were affected, and this led to failure of microspore release. The activity of polygalacturonase and the expression of the pectin methylesterase gene (AT3g06830) were significantly downregulated. Nine genes were downregulated in 7-7365A compared to 7-7365B and 7-7365C, including genes specifically expressed in tapetum (A3, A9, MS1) and the ABA-responsive gene KIN1. ABA concentration in 7-7365B was significantly higher than in 7-7365A and 7-7365C in young flower buds. Furthermore, temperature treatment made some sterile 7-7365A flowers become fertile. The stamens in these flowers produced viable pollen, and filament elongation was restored to its level in 7-7365C. We propose that ABA might control the expression of genes involved in cell separation during early anther development. The REGMS phenotype could be controlled by a primary pathway of male sterile metabolism positively regulated by the BnMs3 gene and a supplementary pathway negatively regulated by the BnRf gene.
Plant Molecular Biology 10/2009; 72(1-2):111-23. · 4.15 Impact Factor
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ABSTRACT: Male sterility in a near-isogenic line S45AB after 25 generations of subcrossing is controlled by two pairs of duplicate genes. The genotype of S45A is Bnms1Bnms1Bnms2Bnms2, and that of S45B is BnMs1Bnms1Bnms2Bnms2, respectively. Histological observations revealed that abnormal anther development appeared in the tapetum and pollen exine during the tetrad stage. This male sterility was characterized by hypertrophy of the tapetal cells at the tetrad stage and a complete lack of microspore exine after the release of microspores from the tetrads. To elucidate the mechanism of this recessive genic male sterility, the flower bud expression profiles of the S45A and S45B lines were analyzed using an Arabidopsis thaliana ATH1 oligonucleotide array. When compared with the S45B line, 69 genes were significantly downregulated, and 46 genes were significantly upregulated in the S45A line. Real-time polymerase chain reaction (PCR) was then used to verify the results of the microarray analysis, and the majority of the downregulated genes in the S45A line were abundantly and specifically expressed in the anther. The results of the real-time PCR suggest that Bnms1 might be involved in the metabolism of lipid/fatty acids, and the homologous mutation of Bnms1 may either block the biosynthesis of sporopollenin or block sporopollenin from being deposited on the microspore surface, thus, preventing pollen exine formation. The role of Bnms1 in the regulatory network of exine formation is also discussed as well.
Plant Cell Reports 07/2009; 28(9):1363-72. · 2.27 Impact Factor
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ABSTRACT: To investigate the long-term outcome and clinical value of utilizing plasma radio frequency ablation under indirect laryngoscope in treatment of recurrence of adenoidal hypertrophy.
Fifty patients with recurrence of adenoidal hypertrophy were subjected to the operation designed by our group.
The period of follow-up was more then 2 years. All patients were free of snore postoperatively; conductive hearing loss was improved as well.
Operation with plasma radiofrequency ablation had advantages of direct and clear view of surgical area, accurate remove of adenoids, non-bleeding, avoiding damage of normal nasopharyngeal structure structures, minimal invasion, high safety, and little complication, in treatment of recurrence of adenoidal hypertrophy.
Lin chuang er bi yan hou ke za zhi = Journal of clinical otorhinolaryngology 01/2009; 22(24):1127-8, 1140.
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ABSTRACT: Brassica napus (AACC, 2n = 38) is a self-compatible amphidiploid plant that arose from the interspecies hybridization of two self-incompatible species, B. rapa (AA, 2n = 20) and B. oleracea (CC, 2n = 18). Self-incompatibility (S) haplotypes in one self-incompatible line and 124 cultivated B. napus lines were detected using S-locus-specific primers, and their relationships with restorer-maintainers were investigated. Two class I (S-I ( SLG ) a and S-I ( SLG ) b) and four class II (S-II ( SLG ) a, S-II ( SLG ) b, S-II ( SP11 ) a and S-II ( SP11 ) b) S haplotypes were observed, of which S-II ( SP11 ) b was newly identified. The nucleotide sequence of SP11 showed little similarity to the reported SP11 alleles. The lines were found to express a total of eleven S genotypes. The self-incompatible line had a specific genotype consisting of S-II ( SP11 ) a, similar to B. rapa S-60, and S-II ( SLG ) a, similar to B. oleracea S-15. Restorers expressed six genotypes: the most common genotype contained S-I ( SLG ) a, similar to B. rapa S-47, and S-II ( SLG ) b, similar to B. oleracea S-15. Maintainers expressed nine genotypes: the predominant genotype was homozygous for two S haplotypes, S-II ( SLG ) a and S-II ( SP11 ) b. One genotype was specific to restorers and four genotypes were specific to maintainers, whereas five genotypes were expressed in both restorers and maintainers. This suggests that there is no definitive correlation between the distribution of S genotypes and restorer-maintainers of self-incompatibility. The finding that restorers and maintainers express unique genotypes, and share some common genotypes, would be valuable for detecting the interaction of S haplotypes in inter- or intra-genomes as well as for developing markers-assisted selection in self-incompatibility hybrid breeding.
Theoretical and Applied Genetics 08/2008; 117(2):171-9. · 3.30 Impact Factor
