| GhCPS基因沉默对棉花幼苗生长和内源激素含量的影响 |
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| 引用本文： | 王丽,穆春,张明才,杜明伟,田晓莉,李召虎.GhCPS基因沉默对棉花幼苗生长和内源激素含量的影响[J].棉花学报,2014,26(3):189-196. |
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| 作者姓名： | 王丽 穆春 张明才 杜明伟 田晓莉 李召虎 |
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| 作者单位： | 1.植物生长调节剂教育部工程研究中心/中国农业大学农学与生物技术学院，北京100193；2.河南师范大学生命科学学院，河南 新乡453007 |
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| 基金项目： | 国家自然科学基金（31271628） |
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| 摘 要： | 利用RT-PCR从棉花中获得493 bp的GhCPS(赤霉素(GA)合成酶基因)片段,经EcoRⅠ/KpnI双酶切后连入pTRV-RNA2质粒,获得了重组载体pTRV-GhCPS;用该载体转化农杆菌后,室内盆栽条件下侵染棉花品种欣抗4的子叶苗。结果表明,病毒诱导沉默GhCPS基因的棉花幼苗中GhCPS的表达量仅为空载体对照的52%。与空载体对照相比,沉默GhCPS的棉花幼苗第1叶和第2叶的内源GA4含量分别降低33%和60%;第2叶ABA含量降低48%,IAA、ZR和iPA含量均无显著变化;株高降低64%。第1叶和第2叶的叶面积分别减小26%和47%,总叶绿素含量分别增加29%和63%,类胡萝卜素含量分别增加27%和46%。第1叶的净光合速率、气孔导度、胞间CO2和蒸腾速率分别增加23%、40%、9%和31%。可见,利用病毒诱导基因沉默技术技术沉默GhCPS后可调控棉花幼苗叶片内源激素平衡,控制棉花幼苗的生长,并提高单位叶面积的光合能力。
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| 关 键 词： | 棉花 病毒诱导的基因沉默 赤霉素 净光合速率 ABA |
| 收稿时间： | 2013-03-31; |
Effect of Silencing GhCPS on the Growth and Endogenous Hormone Content of Cotton Seedlings(Gossypium hirsutum L.) |
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| Wang Li,Mu Chun,Zhang Mingcai,Du Mingwei,Tian Xiaoli,Li Zhaohu.Effect of Silencing GhCPS on the Growth and Endogenous Hormone Content of Cotton Seedlings(Gossypium hirsutum L.)[J].Cotton Science,2014,26(3):189-196. |
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| Authors: | Wang Li Mu Chun Zhang Mingcai Du Mingwei Tian Xiaoli Li Zhaohu |
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| Institution: | 1.Engineering Research Center of Plant Growth Regulator, Ministry of Education / College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China; 2. College of Life Science, Henan Normal University, Xinxiang, Henan 453007, China |
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| Abstract: | Virus-induced gene silencing(VIGS) is a new reverse genetics method used in rapid identification of gene function in plant and has become one of the most widely used plant functional genomics tools. Ent-copalyl diphosphate synthase(CPS) is the first key enzyme involved in gibberellin (GA) biosynthesis. We aimed to elucidate the effect of silencing GhCPS by VIGS on the growth and endogenous hormone content of cotton seedlings. In this study, a 493-bp fragment of GhCPS was amplified from the cotton cultivar(Xinkang 4) cDNA using RT-PCR and was subsequently inserted into the pTRV-RNA2 vector using EcoRⅠand KpnI sites, respectively. pTRV-GhCPS construct was transformed into Agrobacterium tumefaciens GV3101 and was used to transform cotton seedlings at the cotyledon stage with cotton cultivar Xinkang 4 grown in a growth chamber under pot culture. The results showed that the expression level of GhCPS in GhCPS VIGS plants was only 52% of that in pTRV control plants. Silencing GhCPS resulted in 33% and 60% reductions in GA4 content in the 1st and 2nd leaves, respectively, and a slight decrease of IAA content but no obvious changes in ZR and iPA contents in both leaves. ABA content in the 2nd leaf was decreased by 48%. The height of GhCPS VIGS plants was reduced by 64%. The area of the 1st and 2nd leaves in GhCPS VIGS plants was decreased by 26% and 47%, respectively. Silencing GhCPS increased total chlorophyll contents by 29% and 63%, and carotenoids contents by 27% and 46% in the 1st and 2nd leaves, respectively. Net photosynthesis rate, stomatal conductance, intercellular CO2 concentration and transpiration rate in the 1st leaf of GhCPS VIGS plants were enhanced by 23%、40%、9% and 31%, respectively. The results suggest that silencing GhCPS can disturb the balance of endogenous hormone and inhibit the growth of cotton seedlings. The rate of photosynthesis per leaf area can be enhanced by silencing GhCPS. |
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| Keywords: | virus-induced gene silencing gibberellin net photosynthesis rate ABA |
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