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作者： shiraku margaret linyerera 中国农业科学院

学位级别：硕士

棉花是重要的经济作物,也是天然纤维的主要来源。但是,棉花常常受到非生物胁迫（例如干旱、盐和寒冷胁迫）的影响,从而影响了棉花的产量和质量。近年来全球气候变化加剧了非生物胁迫因素的流行和程度。盐和干旱给植物造成渗透胁迫,进而... 详细信息

棉花是重要的经济作物,也是天然纤维的主要来源。但是,棉花常常受到非生物胁迫（例如干旱、盐和寒冷胁迫）的影响,从而影响了棉花的产量和质量。近年来全球气候变化加剧了非生物胁迫因素的流行和程度。盐和干旱给植物造成渗透胁迫,进而对棉花产量产生负面影响。在长期的进化过程中,植物对非生物胁迫产生了适应性应答反应,涉及到植物的生理和代谢途径、转录调控、表观遗传调控,以及在植物发育过程中多基因的表达和相互作用。本研究对LEA3,ACX3和RPL14B等植物逆境应答基因进行了功能表型鉴定,并对其应激反应相关的分子机制进行了初步探讨,主要结果如下:LEA3蛋白在植物生长、发育和非生物胁迫响应中起着重要的作用。我们对第LEA3基因进行了全基因组鉴定,在陆地棉、海岛棉、毛棉、黄褐棉、亚洲棉和雷蒙德氏棉基因组中分别鉴定到13,14,17,16,7和8个LEA3基因,聚类为六个组。表达谱分析显示,陆地棉的LEA3基因在干旱和盐胁迫下表达量更高,特别是在叶和根中对干旱和盐分胁迫表现出更明显的应答反应。通过构建过表达载体和沉默载体分别转化拟南芥和棉花,选择高表达基因Gh＿A08G0694和Gh＿A10G1465进行功能验证。结果发现沉默Gh LEA3（Gh＿A08G0694和Gh＿A10G1465）增加了棉花植物对盐和干旱胁迫的敏感性,并导致ABA和胁迫相关基因的下调表达。Gh LEA3的过表达增强了转基因拟南芥植株的盐、旱胁迫耐受性,且胁迫相关基因上调表达。生化分析表明,沉默Gh LEA3后植株受到了氧化胁迫,而过表达的植株则没有。亚细胞定位预测该类基因定位于细胞核、叶绿体、线粒体和细胞质中。酵母双杂交结果表明,这两个基因与Gh LEA3蛋白同源,且后者与线粒体外膜蛋白孔蛋白、电压依赖性阴离子通道（VDAC1）和甘油醛-3-磷酸脱氢酶（GAPA）互作。本研究表明Gh LEA3基因通过与Gh VDAC1,Gh GAPA和其他调控网络的相互作用来调控干旱和盐分胁迫耐受性,并通过调节应激相关基因的转录而参与信号传导途径。ACX3基因作为脂肪酸参与β-氧化途径的激活剂,在植物生长发育和逆境响应中起着至关重要的作用。ACX3过氧化物酶体通过β-氧化途径导致下游事件级联,应答生化和生理反应的转导。我们在陆地棉、海岛棉、毛棉、黄褐棉以及达尔文氏棉、亚洲棉和雷蒙德氏棉基因组中分别鉴定到20、18、22、23、20、11和9个ACX3蛋白。陆地棉基因表达谱分析表明,逆境胁迫条件下该基因家族在根和叶中被大量诱导。选择Gh＿D01G0186基因通过过表达和VIGS来进行功能初步分析,发现Gh＿D01G0186过表达的植物对胁迫的耐受性增强。胁迫条件下的转基因拟南芥种子比对照的生长速度更快,存活率也更高。相反,棉花Gh＿D01G0186基因的沉默导致胁迫敏感性增加且根系伸长的减少。因此,与ACX3同源的基因增强了棉花植物的胁迫耐受性,并调节其他胁迫响应基因的表达。RPL基因广泛参与植物非生物胁迫应答,然而其响应机制一直没有被揭示。本研究中,我们在陆地棉（AD1）,亚洲棉（A2）和雷蒙德氏棉（D5）三个不同棉种中分别鉴定出26,9,9个RPL14B基因并在盐/干旱胁迫条件下分析了陆地棉中RPL14B基因的差异表达。从中挑选了Gh＿D01G0234基因进行VIGS分析进行功能验证。沉默Gh＿D01G0234对干旱/盐胁迫下棉花幼苗的生长性能有显著影响。基因沉默的棉花和对照植株在形态和生理特性上差异显著,同时VIGS植物中的抗氧化酶水平显着降低,而氧化酶水平显着更高。本研究通过过表达和VIGS技术,对LEA3,ACX3和RPL14B进行功能鉴定,揭示了这些基因在增强棉花盐胁迫和干旱胁迫方面的潜力。对这些基因通过基因工程育种,遗传转化到棉花中,对改良棉花对极端气候的适应能力具有重要的生产意义。

关键词：棉花 LEA ACX RPL 超表达病毒诱导的基因沉默(VIGS) 胁迫应答

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Knockdown of 60S ribosomal protein L14‑2 reveals their potential regulatory roles to enhance drought and salt tolerance in cotton

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Journal of Cotton Research 2021年第4期4卷 308-321页

作者： shiraku margaret linyerera MAGWANGA Richard Odongo CAI Xiaoyan KIRUNGU Joy Nyangasi XU Yanchao MEHARI Teame Gereziher HOU Yuqing WANG Yuhong WANG Kunbo PENG Renhai ZHOU Zhongli LIU Fang State Key Laboratory of Cotton Biology/Institute of Cotton Research Chinese Academy of Agricultural Science(ICRCAAS)Anyang 455000HenanChina School of Biological and Physical Sciences(SBPS) Main CampusJaramogi Oginga Odinga University of Science and Technology(JOOUST)Main CampusP.O.Box 210‑40601BondoKenya Anyang Institute of Technology Anyang 455000HenanChina School of Agricultural Sciences Zhengzhou UniversityZhengzhou 450001HenanChina

Background:Cotton is a valuable economic crop and the main significant source of natural fiber for textile industries *** effects of drought and salt stress pose a challenge to strong fiber and large-scale production due to the ever-changing climatic ***,plants have evolved a number of survival strategies,among them is the induction of various stress-responsive genes such as the ribosomal protein large(RPL)*** RPL gene families encode critical proteins,which alleviate the effects of drought and salt stress in *** this study,comprehensive and functional analysis of the cotton RPL genes was carried out under drought and salt ***:Based on the genome-wide evaluation,26,8,and 5 proteins containing the RPL14B domain were identified in Gossypium hirsutum,***,and ***,***,through bioinformatics analysis,key cis-regulatory elements related to RPL14B genes were *** Myb binding sites(MBS),abscisic acid-responsive element(ABRE),CAAT-box,TATA box,TGACG-motif,and CGTCA-motif responsive to methyl jasmonate,as well as the TCA-motif responsive to salicylic acid,were *** analysis revealed a key gene,Gh_D01G0234(RPL14B),with significantly higher induction levels was further evaluated through a reverse genetic *** knockdown of Gh_D01G0234(RPL14B)significantly affected the performance of cotton seedlings under drought/salt stress conditions,as evidenced by a substantial reduction in various morphological and physiological ***,the level of the antioxidant enzyme was significantly reduced in VIGS-plants,while oxidant enzyme levels increased significantly,as demonstrated by the higher malondialdehyde concentration ***:The results revealed the potential role of the RPL14B gene in promoting the induction of antioxidant enzymes,which are key in oxidizing the various *** key pathways need to be investigated and even as we exploit these genes

关键词： Abiotic stress Cotton Ribosomal protein large Transcription factor Virus-induced gene silencing

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Genome-wide identification and analysis of the CNGC gene family in upland cotton under multiple stress conditions

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Journal of Cotton Research 2023年第3期6卷 195-208页

作者： KIRUNGU Joy Nyangasi MAGWANGA Richard Odongo shiraku margaret linyerera OKUTO Erick CAI Xiaoyan XU Yanchao HOU Yuqing AGONG' Stephen Gaya WANG Kunbo WANG Yuhong ZHOU Zhongli LIU Fang State Key Laboratory of Cotton Biology/Institute of Cotton Research Chinese Academy of Agricultural Science(ICR-CAAS)AnyangHenan 455000China School of Biological PhysicalMathematics and Actuarial Sciences(SBPMAS)Jaramogi Oginga Odinga University of Science and Technology(JOOUST)Bondo210‑40601Kenya School of Agricultural Sciences Zhengzhou UniversityZhengzhouHenan 450001China

Background The cyclic nucleotide-gated channel(CNGC)gene family plays a significant role in the uptake of both essential and toxic cations,and has a role in enhancing tolerance to various forms of abiotic stresses as well as the modulation of the heavy metal toxicity to plant through the absorption of heavy *** A complete genome-wide identification and functional characterization of the cotton CNGC genes was carried out,in which 55,28,and 29 CNGC genes were identified in Gossypium hirsutum,***,and ***,*** protein encoded by the CNGC genes exhibited GRAVY value below zero,indicating their hydrophilic *** genes were unevenly distributed in 19 out of 26 chromosomes,in which the highest density were observed on Ah05,with 8 *** gene coverage was observed among the diploid cotton species,with CNGC genes mapped on all A chromosomes and on 11 out of 13 of D *** majority of CNGC proteins were localized in the endoplasmic reticulum,nucleus,and plasma *** expression analysis revealed the up-regulation of Gh_A01G0520(CNGC4)and Gh_D13G1974(CNGC5)across various forms of abiotic ***,down-regulation of Gh_A01G0520(CNGC4)and Gh_D13G1974(CNGC5)in CNGCs silenced plants caused the significantly reduced ability to tolerate drought and salt *** CNGCs silenced plants were recorded to have significantly low content of antioxidants but relatively higher content of oxidant,including MDA and H_(2)O_(2).Furthermore,SPAD,CMS(cell membrane stability),ELWL(excised leaf water loss),SDW(shoot dry matter weight),and RDW(root dry matter weight)were all lower in CNGCs silenced plants compared with the wild type *** Significant reduction in antioxidant content and negative effects of physiological and morphological characters in CNGCs silenced plants has revealed the novel role of CNGC genes in enhancing cell integrity under abiotic stress *** results provide vital information

关键词： Cyclic nucleotide-gated channel Abiotic stress Differential expression VIGS-plants Chromosome Tetraploid cotton

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Genetic map construction and functional characterization of genes within the segregation distortion regions(SDRs)in the F_(2:3) populations derived from wild cotton species of the D genome

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Journal of Cotton Research 2020年第4期3卷 278-296页

作者： KIRUNGU Joy Nyangasi MAGWANGA Richard Odongo shiraku margaret linyerera LU Pu MEHARI Teame Gereziher XU Yuanchao HOU Yuqing AGONG Stephen Gaya ZHOU Yun CAI Xiaoyan ZHOU Zhongli WANG Kunbo LIU Fang State Key Laboratory of Cotton Biology/Institute of Cotton Research Chinese Academy of Agricultural ScienceAnyang 455000HenanChina School of Biological PhysicalMathematics and Actuarial Sciences(SBPMAS)Jaramogi Oginga Odinga University of Science and Technology(JOOUST)210-40601BondoKenya School of Life Science Henan University/State Key Laboratory of Cotton Biology/Henan Key Laboratory of Plant Stress BiologyKeifang 475004HenanChina School of Agricultural Sciences Zhengzhou UniversityZhengzhou 450001HenanChina.

Background:Segregation distortion(SD)is a common phenomenon among stable or segregating populations,and the principle behind it still puzzles many *** F2:3 progenies developed from the wild cotton species of the D genomes were used to investigate the possible plant transcription factors within the segregation distortion regions(SDRs).A consensus map was developed between two maps from the four D genomes,map A derived from F2:3 progenies of Gossypium klotzschianum and *** while Map B from *** and *** F2:3 *** each map,188 individual plants were ***:The consensus linkage map had 1492 markers across the 13 linkage groups with a map size of 1467.445 cM and an average marker distance of 1.0370 *** D502 had the highest percentage of SD with 58.6%,followed by Chromosome D507 with 47.9%.Six thousand and thirty-eight genes were mined within the SDRs on chromosome D502 and D507 of the consensus *** chromosome D502 and D507,2308 and 3730 genes were mined,respectively,and were found to belong to 1117 gourp out of which 622 groups were common across the two ***,genes within the top 9 groups related to plant resistance genes(R genes),whereas 188 genes encoding protein kinase domain(PF00069)comprised the largest *** analysis of the dominant gene group revealed that 287 miRNAs were found to target various genes,such as the gra-miR398,gramiR5207,miR164a,miR164b,miR164c among others,which have been found to target top-ranked stress-responsive transcription factors such as NAC ***,some of the stress-responsive cis-regulatory elements were also ***,RNA profiling of the genes from the dominant family showed that higher numbers of genes were highly upregulated under salt and osmotic stress conditions,and also they were highly expressed at different stages of fiber ***:The results indicated the critical role of the SDRs in the evolution of the k

关键词： Genetic map Segregation distortion region Cis-regulatory elements Genes miRNA

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