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Inheritance and molecular characterization of resistance to AHAS- inhibiting herbicides in rapeseed

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Journal of Integrative Agriculture 2017年第11期16卷 2421-2433页

作者： HU Mao-long PU Hui-ming GAO Jian-qin LONG Wei-hua CHEN Feng ZHOU Xiao-ying ZHANG Wei PENG Qi CHEN Song ZHANG Jie-fu Nanjing Sub-Center of the National Center Oil Crops Improvement/Key Laboratory of Cotton and Rapeseed of the Ministry of Agriculture/Institute of Industrial Crops Jiangsu Academy of Agricultural Sciences Nanjing 210014 P.R. China

Rapeseed is a very important oil crop in China; however, its production is challenging due to the absence of effective weed management strategies. This is predominantly because of a shortage of herbicide resistance genes. Acetohydroxyacid synthase （AHAS） herbicides inhibit AHAS, a key enzyme involved in branched-chain amino acid synthesis that is required for plant growth. A rapeseed line designated M342 with AHAS herbicide resistance was developed through seed muta- genesis and was studied to assess the level and mode of inheritance of the resistance and to identify the molecular basis of resistance. M342 possessed a high level of cross-resistance to sulfonylureas （SUs） and imidazolinones （IMIs）. This resistance was due to AHAS insensitivity to these herbicides and was inherited as a dominant trait conferred by a single nuclear-encoded gene. Molecular analysis revealed the presence of a Trp574Leu mutation in M342, and an allele-specific cleaved amplified polymorphic sequence （AS-CAPS） marker was developed and cosegregated with herbicide resistance in the F2, BC1, and BC2 populations. This mutation altered the transcript levels of BnAHAS1 and BnAHAS3 in M342 compared with those in the wild type, but it did not affect the agronomic or quality traits. The simple genetic inheritance of this mutation and the availability of the cleaved amplified polymorphic sequence （CAPS） marker and herbicide resistance gene should facilitate the development of herbicide-resistant rapeseed cultivars for effective weed control in China.

关键词： rapeseed acetohydroxyacjd synthase (AHAS) sulfonylurea imidazolinone herbicide resistance mutation

Molecular evidence for blocking erucic acid synthesis in rapeseed(Brassica napus L.) by a two-base-pair deletion in FAE1(fatty acid elongase 1)

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Journal of Integrative Agriculture 2015年第7期14卷 1251-1260页

作者： WU Lei JIA Yan-li WU Gang LU Chang-ming oil crops Research institute Chinese Academy of Agricultural Sciences/Key Laboratory of Biology and Genetic Improvement of Oil CropsMinistry of Agriculture

DNA sequences of fatty acid elongase 1 genes FAE1.1 （EA） and FAE1.2 （Ec） were isolated and characterized for 30 com- mercialized low erucic acid rapeseed （LEAR） cultivars in China. Four types of independent mutation leading to low erucic acid trait were found, i.e., a single-base transition （eAl）, a two-base deletion （ec2） and four-base deletion （eC4） as well as single-base transition with a four-base deletion （eA.）. Three genotypes, i.e., eA1eA1eC2eC2, eA1eA1eC4eC4 and ***4ec4 were responsible for LEA content in storage Iipids of different rapeseed cultivars. Most of the LEAR cultivars had a genotype of eA1eA1ec2ec2, which were descended from the first LEAR cultivar, Oro. Yeast expression analysis revealed that two-base-pair （AA） deletion （ec2） at the base sites of 1 422-1 423 in the C genome FAE1 gene resulted in the absence of the condensing enzyme and led to the failure to produce erucic acid. Coexpression of FAE1 and ketoacyI-CoA reductase （KCR） or enoyI-CoA reductase （ECR） was found in high erucic acid rapeseed （HEAR） but not in LEAR （eA1eA1ec2ec2oreA1eA1ec4ec4）. Moreover, KCR and ECR were still coordinately regulated in eA1eA1ec2ec2 or eA1eA1ec4ec4 genotypes, suggesting that the expression of two genes was tightly linked. In addition, specific detection methods were developed by high-resolution melting curve analysis in order to detect eA1 and ec4.

关键词： erucic acid fatty acid elongase 1 natural mutation Brassica napus L.

Association Analysis for Quality Traits in a Diverse Panel of Chinese Sesame(Sesamum indicum L.)Germplasm

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Journal of Integrative Plant Biology 2013年第8期55卷 745-758页

作者： Wenliang Wei Yanxin Zhang Haixia Lü Donghua Li Linhai Wang Xiurong Zhang oil crops Research institute of the Chinese Academy of Agricultural Sciences Key Laboratory of Biology and Genetic Improvement of Oil CropsMinistry of Agriculture

The main objective of this study was to evaluate the potential of a sesame （Sesamum indicum L.） panel for association analysis, and investigate the genetic basis of oil content （OC）, protein content, oleic acid concentration, and linoleic acid concentration using association mapping. A panel of 216 sesame accessions was phenotyped in a multi-environment trial and fingerprinted with 608 polymorphic loci produced by 79 primers, including simple sequence repeats （SSRs）, sequence-related amplified polymorphisms （SRAPs）, and amplified fragment length polymorphisms （AFLPs）. Population structure analysis revealed two subgroups in the population. The Q model performed better for its ability to re-identify associations for the four traits at highly significant P-values compared to the other three mixed models. And a total of 35 and 25 associations for the four traits in 2010 and 2011 were identified, respectively, with the Q model after Bonferroni correction. Among those associations, only one for OC was re-identified in two environments, and several markers associated simultaneously with multiple traits were discovered. These results suggest the power and stability of the Q model for association analysis of nutritional traits in this sesame panel for its slight population stratification and familial relationship, which could aid in dissecting complex traits, and could help to develop strategies for improving nutritional quality.

关键词： Association mapping nutritional trait population structure Sesamum indicum L.

Comparison of transcriptomes undergoing waterlogging at the seedling stage between tolerant and sensitive varieties of Brassica napus L.

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Journal of Integrative Agriculture 2015年第9期14卷 1723-1734页

作者： ZOU Xi-ling ZENG Liu LU Guang-yuan CHENG Yong XU Jin-song ZHANG Xue-kun Key Laboratory of Biology and Genetic Improvement of oil crops Ministry of Agriculture/Oil Crops Research InstituteChinese Academy of Agricultural Sciences

RNA sequencing of the sensitive GH01 variety of Brassica napus L. seedling roots under 12 h of waterlogging was compared with previously published data of the ZS9 tolerant variety to unravel genetic mechanisms of waterlogging tolerance beyond natural variation. A total of 2 977 genes with similar expression patterns and 17 genes with opposite expression patterns were identiifed in the transcription proifles of ZS9 and GH01. An additional 1 438 genes in ZS9 and 1 861 genes in GH01 showed strain speciifc regulation. Analysis of the overlapped genes between ZS9 and GH01 revealed that waterlogging tolerance is determined by ability to regulate genes with similar expression patterns. Moreover, differences in both gene expression proifles and abscisic acid (ABA) contents between the two varieties suggest that ABA may play some role in waterlogging tolerance. This study identiifes a subset of candidate genes for further functional analysis.

关键词： rapeseed(Brassica napus L.) waterlogging digital gene expression(DGE) roots transcriptome comparative analysis

Genetic dissection of root morphological traits as related to potassium use efficiency in rapeseed under two contrasting potassium levels by hydroponics

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Science China(Life Sciences) 2019年第6期62卷 746-757页

作者： Xiaoling Dun Jiaqin Shi Huiping Liu Jie Wang Xinfa Wang Hanzhong Wang oil crops Research institute of the Chinese Academy of Agricultural Sciences/Key Laboratory of Biology and Genetic Improvement of oil crops Ministry of Agriculture

To reveal the genetic basis of potassium use efficiency(KUE) in rapeseed, root morphology(RM), biomass and KUE-related traits were measured in a recombinant inbred line population with 175 F7 lines that were subjected to high-potassium(HK) and low-potassium(LK) treatments by hydroponics. A total of 109 significant QTLs were identified to be associated with the examined traits. Sixty-one of these QTLs were integrated into nine stable QTLs. The higher heritability for RM and biomass traits and lower heritability for KUE-related traits, as well as nine stable QTLs for RM traits and only two for KUE-related traits,suggested that regulating RM traits would be more effective than selecting KUE traits directly to improve KUE by markerassisted selection. Furthermore, the integration of stable QTLs identified in the HK, LK, high-nitrogen(HN) and low-nitrogen(LN) conditions gave 10 QTL clusters. Seven of these clusters were classified into major QTLs that explained 7.4%–23.7% of the total phenotypic variation. Five of the major QTL clusters were detected under all of the treated conditions, and four clusters were specifically detected under the LK and LN conditions. These common and specific QTL clusters may be useful for the simultaneous improvement of multiple traits by marker-assisted selection.

关键词： Brassica napus L. root traits potassium efficiency QTL mapping QTL cluster

Detection of DNA methylation changes during seed germination in rapeseed (Brassica napus)

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Chinese Science Bulletin 2006年第2期51卷 182-190页

作者： LU Guangyuan WU Xiaoming CHEN Biyun GAO Guizhen XU Kun LI Xiangzhi Key Laboratory of oil crops Genetic Improvement of the Ministry of Agriculture Oil Crops Research Institute CAAS Wuhan 430062 China

DNA methylation is known to play a crucial role in regulating plant development and or- gan or tissue differentiation. In this study, we as- sessed the extent and pattern of cytosine methylation during rapeseed (Brassica napus L.) seed germina- tion, and compared the methylation level of various tissues in seedling, using the techniques of methyla- tion-sensitive amplified polymorphism (MSAP) and HPLC separation and quantification of nucleosides. In all, 484 bands, each representing a recognition site cleaved by either or both of the isoschizomers, were amplified by 12 pairs of selective primers in DNA ob- tained from dry seeds. A total of 76 sites were found to be differentially digested by the isoschizomers, indicating that approximately 15.7% of 5′-CCGG-3′ sites in the genome were cytosine methylated. Four classes of patterns were observed in a comparative assay of cytosine methylation in the dry and germi- nating seeds; a small number of hypermethylation events occurred at 5′-CCGG-3′ sites in germinating seeds compared with dry seeds, while many more hypomethylation events were detected after seed germination. Differences in DNA methylation level in various tissues were also detected; radicel was less methylated than hypocotyl and cotyledon. These observations were further confirmed by HPLC analy- sis. In addition, sequencing of eleven differentially methylated fragments and the subsequent blast search revealed that cytosine methylated 5′-CCGG- 3′ sequences were equally distributed between cod- ing and non-coding regions. These results clearly demonstrate the power of MSAP technique for large-scale DNA methylation detection in rapeseed genome, and the complexity of DNA methylationchange during seed germination. DNA Hypomethyla- tion going with seed germination appears to be a necessary step toward transcriptional activation in gene expression, and may well contribute to the de- velopmental gene regulation.

关键词：油菜籽芸苔属 DNA甲基化作用种子萌芽 MSAP HPLC

Diversity characterization and association analysis of agronomic traits in a Chinese peanut (Arachis hypogaea L.) mini-core collection

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Journal of Integrative Plant Biology 2014年第2期56卷 159-169页

作者： Huifang Jiang Li Huang Xiaoping Ren Yuning Chen Xiaojing Zhou Youlin Xia Jiaquan Huang Yong Lei Liying Yan Liyun Wan Boshou Liao Key Laboratory of Biology and Genetic Improvement of oil crops Ministry of Agriculture Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences Nanchong Academy of Agricultural Sciences

Association mapping is a powerful approach for exploring the molecular basis of phenotypic variations in plants. A peanut （Arachis hypogaea L.） mini-core collection in China comprising 298 accessions was genotyped using lo9 simple sequence repeat （SSR） markers, which identified 554 SSR alleles and phenotyped for 15 agronomic traits in three different environments, exhibiting abundant genetic and phenotypic diversity within the panel. A model-based structure analysis assigned all accessions to three groups. Most of the accessions had the relative kinship of less than o.05, indicating that there were no or weak relationships between accessions of the mini- core collection. For 15 agronomic traits in the peanut panel, generally the Q ＋ K model exhibited the best performance to eliminate the false associated positives compared to the Q model and the general linear model-simple model. In total, 89 SSR alleles were identified to be associated with 15 agronomic traits of three environments by the Q＋K model-based association analysis. Of these, eight alleles were repeatedly detected in two or three environments, and 15 alleles were commonly detected to be associated with multiple agronomic traits. Simple sequence repeat allelic effects confirmed significant differences between different genotypes of these repeatedly detected markers. Our results demonstrate the great potential of integrating the association analysis and marker-assisted breeding by utilizing the peanut mini-core collection.

关键词： Arachis hypogaea L. association analysis mini-corecollection peanut

Rapeseed research and production in China

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The Crop Journal 2017年第2期5卷 127-135页

作者： Qiong Hu Wei Hua Yan Yin Xuekun Zhang Lijiang Liu Jiaqin Shi Yongguo Zhao Lu Qin Chang Chen Hanzhong Wang oil crops Research institute Chinese Academy of Agricultural Sciences/Key Laboratory for Biological Sciences and Genetic Improvement of Oil Crops Ministry of Agriculture Wuhan 430062 China

Rapeseed(Brassica napus L.) is the largest oilseed crop in China and accounts for about 20% of world *** the last 10 years,the production,planting area,and yield of rapeseed have been stable,with improvement of seed quality and especially seed oil *** is among the leading countries in rapeseed genomic research internationally,having jointly with other countries accomplished the whole genome sequencing of rapeseed and its two parental species,Brassica oleracea and Brassica *** on functional genomics including the identification of QTL governing important agronomic traits such as yield,seed oil content,fertility regulation,disease and insect resistance,abiotic stress,nutrition use efficiency,and pod shattering resistance has been *** a consequence,molecular markers have been developed and used in breeding *** 2005–2014,215 rapeseed varieties were registered nationally,including 210 winter-and 5 spring-type *** across the whole process of rapeseed production was investigated and operating instructions for all relevant techniques were *** techniques for rapeseed field management such as high-density planting,controlled-release fertilizer,and biocontrol of disease and pests combined with precision tools such as drones have been developed and are being adopted in *** the application of advanced breeding and production technologies,in the near future,the oil yield and quality of rapeseed varieties will be greatly increased,and more varieties with desirable traits,especially early maturation,high yield,high resistance to biotic and abiotic stress,and suitability for mechanized harvesting will be *** of modern technologies on the mechanized management of rapeseed will greatly increase grower profit.

关键词： Rapeseed Review Genomics Breeding Cultivation Mechanization

Laboratory testing and molecular analysis of the resistance of wild and cultivated soybeans to cotton bollworm, Helicoverpa armigera(Hübner)

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The Crop Journal 2015年第1期3卷 19-28页

作者： Xiaoyi Wang Haifeng Chen Aihua Sha Rong Zhou Zhihui Shang Xiaojuan Zhang Chanjuan Zhang Limiao Chen Qingnan Hao Zhonglu Yang Dezhen Qiu Shuilian Chen Xinan Zhou Department of Soybean Oil Crops Research Institute Chinese Academy of Agricultural Sciences

Identifying a superior soybean variety with high defoliator resistance is important to avoid yield loss. cotton bollworm(Helicoverpa armigera Hübner) is one of the major defoliators of soybean(Glycine max [L.] Merr.) worldwide. In this study, we evaluated the effect of H. armigera larvae on ED059, a wild soybean(Glycine soja Sieb. et Zucc.), and three cultivated soybean varieties: Tianlong 2, PI 535807, and PI 533604, in choice and no-choice assays. The percentage of ED059 leaflets consumed by H. armigera was lower than that of the three cultivated soybeans. Larvae that fed on ED059 exhibited low weight gain and high mortality *** nutritional indices suggested that ED059 reduced the growth, consumption, and frass production of H. armigera larvae. Larvae that fed on ED059 showed lower efficiency of conversion of ingested and of digested food than those that fed on Tianlong 2 and PI ***, they showed statistically similar consumption index and approximate digestibility compared with those fed on the three cultivated soybeans. Quantitative real-time PCR analysis revealed that 24 h after insect attack, ED059 had higher transcript levels of Kunitz trypsin inhibitor 3, Cysteine proteinase inhibitor 2, and Nerolidol synthase 1 but a lower transcript level of Pathogenesis-related protein 1 than Tianlong 2. The gene expression results were consistent with the presence of higher levels of jasmonic acid(JA) and transcript levels of the JA biosynthesis enzyme allene oxide cyclase 3 in ED059 than in Tianlong 2. Our findings indicate that ED059 is a superior soybean line with strong insect resistance that may be mediated via the JA pathway.

关键词： Soybean Defoliation Defoliator resistance Real-time PCR