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Pyramiding of multiple genes generates rapeseed introgression lines with clubroot and herbicide resistance, high oleic acid content, and early maturity

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The Crop Journal 2023年第3期11卷 895-903页

作者： Zhaoyang Wang Fucai Wang Zihan Yu Xiaorui Shi Xianming Zhou Pengfei Wang Yixian Song Dengfeng Hong Guangsheng Yang National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural UniversityWuhan 430070HubeiChina Hubei Hongshan Laboratory Wuhan 430070HubeiChina Sanya Nanfan Research Institute of Hainan University Hainan Yazhou Bay Seed LaboratorySanya 572025HainanChina College of Tropical Crops Hainan University Hainan UniversityHaikou 570288HainanChina

Clubroot and herbicide resistance,high oleic acid(OA)content,and early maturity are targets of rapeseed(Brassica napus L.)*** objective of this study was to develop new male-fertility restorer lines by pyramiding favorable genes to improve these traits *** elite alleles for the four traits were introduced into the restorer line 621R by speed breeding with marker-assisted and phenotypic *** introgression lines(ILs)were developed with four-to seven-gene combinations and crossed with two elite parents to develop *** ILs and their corresponding hybrids displayed high resistance to both clubroot pathotype 4 and sulfonylurea *** ILs and their hybrids showed large increases in OA contents and four showed earlier *** new ILs may be useful in rapeseed hybrid breeding for the target traits.

关键词： Clubroot resistance herbicide resistance Oleic acid content Flowering time Pyramid breeding

Genomic insights into the origin, adaptive evolution, and herbicide resistance of Leptochloa chinensis, a devastating tetraploid weedy grass in rice fields

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Molecular Plant 2022年第6期15卷 1045-1058页

作者： Lifeng Wang Xuepeng Sun Yajun Peng Ke Chen Shan Wu Yanan Guo Jingyuan Zhang Haona Yang Tao Jin Lamei Wu Xiaomao Zhou Bin Liang Zhenghong Zhao Ducai Liu Zhangjun Fei Lianyang Bai State Key Laboratory of Hybrid Rice Hunan Academy of Agricultural SciencesChangsha 410125China Hunan Weed Science Key Laboratory Hunan Academy of Agricultural SciencesChangsha 410125China Boyce Thompson Institute Cornell UniversityIthacaNY 14853USA College of Horticulture Science Zhejiang A&F UniversityHangzhou 311300China Qingdao Kingagroot Compounds Co.Ltd Qingdao 266000China USDA-ARS Robert W.Holley Center for Agriculture and HealthIthacaNY 14853USA

Chinese sprangletop (Leptochloa chinensis), belonging to the grass subfamily Chloridoideae, is one of the most notorious weeds in rice ecosystems. Here, we report a chromosome-scale reference genome assembly and a genomic variation map of the tetraploid L. chinensis. The L. chinensis genome is derived from two diploid progenitors that diverged ∼10.9 million years ago, and its two subgenomes display neither fractionation bias nor overall gene expression dominance. Comparative genomic analyses reveal substantial genome rearrangements in L. chinensis after its divergence from the common ancestor of Chloridoideae and, together with transcriptome profiling, demonstrate the important contribution of tetraploidization to the gene sources for the herbicide resistance of L. chinensis. Population genomic analyses of 89 accessions from China reveal that L. chinensis accessions collected from southern/southwestern provinces have substantially higher nucleotide diversity than those from the middle and lower reaches of the Yangtze River, suggesting that L. chinensis spread in China from the southern/southwestern provinces to the middle and lower reaches of the Yangtze River. During this spread, L. chinensis developed significantly increased herbicide resistance, accompanied by the selection of numerous genes involved in herbicide resistance. Taken together, our study generated valuable genomic resources for future fundamental research and agricultural management of L. chinensis, and provides significant new insights into the herbicide resistance as well as the origin and adaptive evolution of L. chinensis.

关键词： Leptochloa weed sgenome polyploidization genome evolution genetic diversity adaptive selection herbicide resistance

Experimental and computational correlation and prediction on herbicide resistance for acetohydroxyacid synthase mutants to Bispyribac

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Science China Chemistry 2013年第3期56卷 286-405页

作者： HE YinWu NIU CongWei LI Hank WEN Xin XI Zhen State Key Laboratory of Elemento-Organic Chemistry

Bispyribac is a widely used herbicide that targets the acetohydroxyacid synthase(AHAS) *** in AHAS have caused serious herbicide resistance that threatened the continued use of the *** far,a unified model to decipher herbicide resistance in molecular level with good prediction is still *** this paper,we have established a new QSAR method to construct a prediction model for AHAS mutation resistance to herbicide Bispyribac.A series of AHAS mutants concerned with the herbicide resistance were constructed,and the inhibitory properties of Bispyribac against these mutants were *** 3D-QSAR method has been transformed to process the AHAS mutants and proposed as mutation-dependent biomacromolecular QSAR(MB-QSAR).The excellent correlation between experimental and computational data gave the MB-QSAR/CoMFA model(q2=0.615,r2=0.921,r2 pred=0.598) and the MB-QSAR/CoMSIA model(q2=0.446,r2=0.929,r2 pred=0.612),which showed good prediction for the inhibition properties of Bispyribac against AHAS *** MB-QSAR models,containing the three-dimensional molecular interaction diagram,not only disclose to us for the first time the detailed three-dimensional information about the structure-resistance relationships,but may also provide further guidance to resistance mutation ***,the molecular interaction diagram derived from MB-QSAR models may aid the resistance-evading herbicide design.

关键词： MB-QSAR resistance prediction herbicide resistance Bispyribac-sodium acetohydroxyacid synthase

Experimental and computational correlation and prediction on herbicide resistance for acetohydroxyacid synthase mutants to Bispyribac

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Science China(Physics,Mechanics & Astronomy) 2013年第3期56卷 286-295页

Bispyribac is a widely used herbicide that targets the acetohydroxyacid synthase （AHAS） enzyme. Mutations in AHAS have caused serious herbicide resistance that threatened the continued use of the herbicide. So far, a unified model to decipher herb- icide resistance in molecular level with good prediction is still lacking. In this paper, we have established a new QSAR method to construct a prediction model for AHAS mutation resistance to herbicide Bispyribac. A series of AHAS mutants concerned with the herbicide resistance were constructed, and the inhibitory properties of Bispyribac against these mutants were meas- ured. The 3D-QSAR method has been transformed to process the AHAS mutants and proposed as mutation-dependent biom- acromolecular QSAR （MB-QSAR）. The excellent correlation between experimental and computational data gave the MB-QSAR/CoMFA model （q2 = 0.615, P = 0.921, F2pred = 0.598） and the MB-QSAR/CoMSIA model （q2 = 0.446, r2 = 0.929, r2pred = 0.612）, which showed good prediction for the inhibition properties of Bispyribac against AHAS mutants. Such MB-QSAR models, containing the three-dimensional molecular interaction diagram, not only disclose to us for the first time the detailed three-dimensional information about the structure-resistance relationships, but may also provide further guidance to resistance mutation evolution. Also, the molecular interaction diagram derived from MB-QSAR models may aid the resistance-evading herbicide design.

关键词： MB-QSAR resistance prediction herbicide resistance Bispyribac-sodium acetohydroxyacid synthase

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

Alleles contributing to acetyl coenzyme A carboxylase(ACCase)resistance in keng stiffgrass(Pseudosclerochloa kengiana)populations from China

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Journal of Integrative Agriculture 2017年第1期16卷 125-134页

作者： YUAN Guo-hui GUO Wen-lei DU Long LIU Wei-tang LI Qi LI Ling-xu WANG Jin-xin College of Plant Protection Shandong Agricultural University College of Chemistry and Pharmacy Science Qingdao Agricultural University

Keng stiffgrass is a grass weed that affects wheat-rice cropping systems in China. The extensive reliance on acetyl coen- zyme A carboxylase （ACCase）-inhibiting herbicides has resulted in keng stiffgrass developing resistance to these herbicides. The objective of this research was to evaluate the resistance level of the putative resistant keng stiffgrass populations to ACCase-inhibiting herbicides and to identify their molecular resistance mechanism. Whole-plant dose-response experi- ments demonstrated that SD-4 （R）, SD-11 （R）, and JS-25 （R） populations were highly resistant to fenoxaprop, clodinafop, and fluazifop, moderately resistant to diclofop, had low resistance to sethoxydim and pinoxaden, but were sensitive to clethodim. Partial chloroplasticACCase sequences showed that there were two copies ofACCase gene in keng stiffgrass, and all homoeologous genes were expressed. The results of sequence analyses of the ACCase CT domain revealed an isoleucine-to-asparagine substitution at position 2041 in SD-4 （R） and SD-11 （R） populations, and a tryptophan-to-cysteine substitution at position 2027 in the JS-25 （R） population. To our knowledge, this is the first report of Ile-2041-Asn and Trp- 2027-Cys mutations in ACCase-resistant keng stiffgrass. In addition, three robust （derived） cleaved amplified polymorphic sequence （（d）CAPS） markers have been developed to rapidly identify these mutations in the ACCase gene of keng stiffgrass.

关键词： herbicide resistance ACCase gene mutation (d)CAPS

Transgenic rice expressing a novel phytase-lactoferricin fusion gene to improve phosphorus availability and antibacterial activity

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Journal of Integrative Agriculture 2017年第4期16卷 774-788页

作者： WANG Zuo-ping DENG Li-hua WENG Lü-shui DENG Xiang-yang FU Xi-qin XIN Ye-yun XIAO Guo-ying Key Laboratory of Agro-ecological Processes in Subtropical Region Institute of Subtropical Agriculture Chinese Academy of Sciences Changsha 410125 P.R.China Graduate University of Chinese Academy of Sciences Beijing 100049 P.R.China State Key Laboratory of Hybrid Rice Hunan Hybrid Rice Research Center Changsha 410125 P.R.China

The developing trends of livestock production are efficiency availability of phytate phosphorus and abuse of antibiotics. safety and sustainability, which face two major challenges： low As a solution phytases and antimicrobial peptides are applied as feed additives. However, phytases and antimicrobial peptides are susceptible to proteases, costly by fermentation and potential toxic to production hosts. We transformed an optimized phytase-lactoferricin fusion gene PhyLfdriven by an en- dosperm-specific promoter Gt13aP and Bar （bialaphos resistance） gene as a selection maker into rice. The Bar and PhyLf genes were integrated into the rice genome, stably inherited and expressed. Their phosphinothricin acetyl transferase （PAT） protein content oftransgenic plants with glufosinate resistance varied between 50.45-93.39 IJg g-l. Fusion protein expressed especially in the seeds of transgenic rice had a summit phytase activity at 32.30 U g-l, which increased by 61.71-fold com- pared to the control/check group （CK） and 7.54-fold compared to un-optimized transgenic plant. The highest inorganic phosphorus （Pi） content of the transgenic seeds reached 13.15 mg g-~, increased by 12.77-fold compared to that of CK. Preliminary antibacterial experiments showed that the enterokinase hydrolysate product of fusion protein could inhibit the growth of Escherichia coil DH5a. These results indicated that the protein PhyLf has the potential to increase availability of feed phytate phosphorus, improve consumer＇s immunity and reduce the use of antibiotics.

关键词： rice phytase antimicrobial peptide lactoferricin fusion gene Bar gene herbicide resistance

EPSPS regulates cell elongation by disrupting the balance of lignin and flavonoid biosynthesis in cotton

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Journal of Integrative Agriculture 2024年第10期23卷 3437-3456页

作者： Qingdi Yan Wei Hu Chenxu Gao Lan Yang Jiaxian Yang Renju Liu Masum Billah Yongjun Lin Ji Liu Pengfei Miao Zhaoen Yang Fuguang Li Wenqiang Qin State Key Laboratory of Cotton Bio‑breeding and Integrated Utilization/Institute of Cotton Research Chinese Academy ofAgricultural SciencesAnyang 455000China National Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research Huazhong AgriculturalUniversityWuhan 430070China State Key Laboratory of Cotton Bio-breeding and Integrated Utilization/Zhengzhou University Zhengzhou 450066China National Nanfan Research Institute(Sanya) Chinese Academy of Agricultural SciencesSanya 572025China

EPSPS is a key gene in the shikimic acid synthesis pathway that has been widely used in breeding crops with herbicide ***,its role in regulating cell elongation is poorly *** the overexpression of EPSPS genes,we generated lines resistant to glyphosate that exhibit an unexpected dwarf phenotype.A representative line,DHR1,exhibits a stable dwarf phenotype throughout its entire growth *** for plant height,the other agronomic traits of DHR1 are similar to its transgenic explants *** section observations showed that DHR1 internodes are shortened due to reduced elongation and division of the internode *** hormones confirmed that DHR1 is not a classical brassinolide(BR)-or gibberellin(GA)-related dwarfing *** analysis and fine mapping confirmed that the EPSPS gene is the causal gene for dwarfism,and the phenotype can be inherited in different *** and metabolome analyses showed that genes associated with the phenylpropanoid synthesis pathway are enriched in DHR1 compared with *** metabolites are enriched in DHR1,whereas lignin metabolites are *** enhancement of flavonoids likely results in differential expression of auxin signal pathway genes and alters the auxin response,subsequently affecting cell *** study provides a new strategy for generating dwarfs and will accelerate advancements in light simplification in the cultivation and mechanized harvesting of cotton.

关键词： EPSPS cotton T-DNA dwarfism herbicide resistance

Scarabaeid Larvae- and herbicide-Resistant Transgenic Perennial Ryegrass (Lolium perenne L.) Obtained by Agrobacterium tumefaciens-Mediated Transformation of cry8Ca2, cry8Ga and bar Genes

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Journal of Integrative Agriculture 2012年第1期11卷 53-61页

作者： WU Jin-xia ZHANG Zhi-guo ZHANG Qian LANG Zhi-hong SUN Xue-hui Biotechnology Research Institute National Key Facility for Crop Gene Resources and Genetic ImprovementChinese Academy of Agricultural SciencesBeijing 100081P.R.China

Insect pest and weeds are two major problems for forage and turf grasses. In this study, scarab larvae- and herbicide-resistant transgenic perennial ryegrass （Lolium perenne L.） was obtained by transforming it with cry and bar genes simultaneously via the Agrobacterium-mediated method. To optimize the callus induction and plant regeneration conditions, various concentrations of 2,4-dichlorophenoxyacetic acid and 6-benzylaminopurine were assayed. The transformation efficiencies of different Agrobacterium suspension media, used during Agrobacterium-mediated transformation, were compared. Then, plasmids of pCAMBIA3301 containing cry gene （cry8Ca2 or cry8Ga） and bar gene, driven by ubiquitin promoter, were transformed into perennial ryegrass. The transformants were generated and confirmed by both Southern hybridization analysis and Western hybridization analysis. Further, the resistance of transgenic perennial ryegrass plants to scarab larvae and herbicide were analyzed. After 30 d of co-cultivation with scarab larvae, the damage to the root system of transgenic plants was less than that of non-transgenic control plants. Additionally, the leaves of transgenic plants were resistant to Basta, while leaves of the wild plants wilted after Basta spraying. These results show that cry gene and bar gene were successfully transferred into perennial ryegrass by the Agrobactgerium-mediated method, and convey resistance to scarab larvae and herbicide in transgenic perennial ryegrass plants.

关键词： Agrobacterium tumefaciens-mediated transformation transgenic perennial ryegrass scarab larvae resistance herbicide resistance