Gh_A08G1120（GH3.5）基因的功能表征揭示了它们在增强棉花的干旱和盐胁迫耐受性中的重要作用。,BMC Genetics

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Gh_A08G1120（GH3.5）基因的功能表征揭示了它们在增强棉花的干旱和盐胁迫耐受性中的重要作用。
BMC Genetics Pub Date : 2019-07-23 , DOI: 10.1186/s12863-019-0756-6
Joy Nyangasi Kirungu ₁ , Richard Odongo Magwanga _{1,

2} , Pu Lu ₁ , Xiaoyan Cai ₁ , Zhongli Zhou ₁ , Xingxing Wang ₁ , Renhai Peng ₃ , Kunbo Wang ₁ , Fang Liu ₁

Affiliation

背景技术生长素在植物生长和发育中起重要作用。生长素反应基因；生长素/吲哚-3-乙酸（Aux / IAA），小生长素上调RNA（SAUR）和Gretchen Hagen3（GH3）控制它们的机制。GH3基因通过生长素结合的催化活性并使游离的吲哚-3-乙酸（IAA）与氨基酸结合，从而在体内稳态中发挥作用。结果在我们的研究中，我们鉴定了三种棉花中的GH3基因。陆地棉，植物棉和雷蒙氏棉，分析了它们的染色体分布，系统发育关系，顺式调控元件功能，并对新的Gh_A08G1120（GH3.5）基因进行了病毒诱导的基因沉默。系统发育树显示了具有进化枝1的四个基因簇在图3和图4中，主要具有棉种GH3的成员，而进化枝2主要是属于拟南芥的成员。没有旁系基因，在棉属和其他物种之间没有观察到直系同源基因。检测到所有GO术语，但发现只有14个基因描述了陆地棉中的GO术语，与其他功能相比，发现了更多的生物学功能。GH3.17亚家族具有最多的顺式调控元件，大多数具有促进脱水反应的启动子。RNA表达分析表明，干旱和盐胁迫条件下的10个基因和8个基因在陆地棉中均被上调。在盐胁迫条件下植物中所有上调的基因在干旱胁迫中也上调。此外，Gh_A08G1120（GH3。5）在两个压力因子上均表现出明显的上调。通过病毒诱导的基因沉默（VIGS）对Gh_A08G1120（GH3.5）的功能表征显示，与野生型相比，VIGS植物耐受干旱和盐胁迫的能力显着降低。在干旱和盐胁迫条件下，VIGS植物中的叶绿素含量，相对叶含水量（RLWC）和超氧化物歧化酶（SOD）浓度水平显着降低，而丙二醛浓度和离子泄漏（作为衡量细胞膜稳定性（CMS）的量度）增加。结论该研究揭示了GH3基因在使植物适应干旱和盐胁迫条件中的重要性，如VIGS结果和RT-qPCR分析所证明的。通过病毒诱导的基因沉默（VIGS）对Gh_A08G1120（GH3.5）的功能表征显示，与野生型相比，VIGS植物耐受干旱和盐胁迫的能力显着降低。在干旱和盐胁迫条件下，VIGS植物中的叶绿素含量，相对叶含水量（RLWC）和超氧化物歧化酶（SOD）浓度水平显着降低，而丙二醛浓度和离子泄漏（作为衡量细胞膜稳定性（CMS）的量度）增加。结论该研究揭示了GH3基因在使植物适应干旱和盐胁迫条件中的重要性，如VIGS结果和RT-qPCR分析所证明的。通过病毒诱导的基因沉默（VIGS）对Gh_A08G1120（GH3.5）的功能表征显示，与野生型相比，VIGS植物耐受干旱和盐胁迫的能力显着降低。在干旱和盐胁迫条件下，VIGS植物中的叶绿素含量，相对叶含水量（RLWC）和超氧化物歧化酶（SOD）浓度水平显着降低，而丙二醛浓度和离子泄漏（作为衡量细胞膜稳定性（CMS）的量度）增加。结论该研究揭示了GH3基因在使植物适应干旱和盐胁迫条件中的重要性，如VIGS结果和RT-qPCR分析所证明的。5）通过病毒诱导的基因沉默（VIGS）显示，与野生型相比，VIGS植物耐受干旱和盐胁迫的能力显着降低。在干旱和盐胁迫条件下，VIGS植物中的叶绿素含量，相对叶含水量（RLWC）和超氧化物歧化酶（SOD）浓度水平显着降低，而丙二醛浓度和离子泄漏（作为衡量细胞膜稳定性（CMS）的量度）增加。结论该研究揭示了GH3基因在使植物适应干旱和盐胁迫条件中的重要性，如VIGS结果和RT-qPCR分析所证明的。5）通过病毒诱导的基因沉默（VIGS）显示，与野生型相比，VIGS植物耐受干旱和盐胁迫的能力显着降低。在干旱和盐胁迫条件下，VIGS植物中的叶绿素含量，相对叶含水量（RLWC）和超氧化物歧化酶（SOD）浓度水平显着降低，而丙二醛浓度和离子泄漏（作为衡量细胞膜稳定性（CMS）的量度）增加。结论该研究揭示了GH3基因在使植物适应干旱和盐胁迫条件中的重要性，如VIGS结果和RT-qPCR分析所证明的。干旱和盐胁迫条件下，VIGS植物中的超氧化物歧化酶和超氧化物歧化酶（SOD）浓度水平显着降低，而丙二醛浓度和离子泄漏（作为衡量细胞膜稳定性（CMS）的量度）增加。结论该研究揭示了GH3基因在使植物适应干旱和盐胁迫条件中的重要性，如VIGS结果和RT-qPCR分析所证明的。在干旱和盐胁迫条件下，VIGS植物中丙二醛和超氧化物歧化酶（SOD）的浓度水平显着降低，而丙二醛浓度和离子泄漏（作为衡量细胞膜稳定性（CMS）的量度）增加。结论该研究揭示了GH3基因在使植物适应干旱和盐胁迫条件中的重要性，如VIGS结果和RT-qPCR分析所证明的。

"点击查看英文标题和摘要"

Functional characterization of Gh_A08G1120 (GH3.5) gene reveal their significant role in enhancing drought and salt stress tolerance in cotton.

BACKGROUND Auxins play an important role in plant growth and development; the auxins responsive gene; auxin/indole-3-acetic acid (Aux/IAA), small auxin-up RNAs (SAUR) and Gretchen Hagen3 (GH3) control their mechanisms. The GH3 genes function in homeostasis by the catalytic activities in auxin conjugation and bounding free indole-3-acetic acid (IAA) to amino acids. RESULTS In our study, we identified the GH3 genes in three cotton species; Gossypium hirsutum, Gossypium arboreum and Gossypium raimondii, analyzed their chromosomal distribution, phylogenetic relationships, cis-regulatory element function and performed virus induced gene silencing of the novel Gh_A08G1120 (GH3.5) gene. The phylogenetic tree showed four clusters of genes with clade 1, 3 and 4 having mainly members of the GH3 of the cotton species while clade 2 was mainly members belonging to Arabidopsis. There were no paralogous genes, and few orthologous genes were observed between Gossypium and other species. All the GO terms were detected, but only 14 genes were found to have described GO terms in upland cotton, more biological functions were detected, as compared to the other functions. The GH3.17 subfamily harbored the highest number of the cis-regulatory elements, most having promoters towards dehydration-responsiveness. The RNA expression analysis revealed that 10 and 8 genes in drought and salinity stress conditions respectively were upregulated in G. hirsutum. All the genes that were upregulated in plants under salt stress conditions were also upregulated in drought stress; moreover, Gh_A08G1120 (GH3.5) exhibited a significant upregulation across the two stress factors. Functional characterization of Gh_A08G1120 (GH3.5) through virus-induced gene silencing (VIGS) revealed that the VIGS plants ability to tolerate drought and salt stresses was significantly reduced compared to the wild types. The chlorophyll content, relative leaf water content (RLWC), and superoxide dismutase (SOD) concentration level were reduced significantly while malondialdehyde concentration and ion leakage as a measure of cell membrane stability (CMS) increased in VIGS plants under drought and salt stress conditions. CONCLUSION This study revealed the significance of the GH3 genes in enabling the plant's adaptation to drought and salt stress conditions as evidenced by the VIGS results and RT-qPCR analysis.

更新日期：2019-07-23

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