Development of Agrobacterium-mediated virus-induced gene silencing and performance evaluation of four marker genes in Gossypium barbadense.
Development of Agrobacterium-mediated virus-induced gene silencing and performance evaluation of four marker genes in Gossypium barbadense.
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DOI：
10.1371/journal.pone.0073211
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Wu J
中科院分区：
文献类型：
--
作者：
Pang J;Zhu Y;Li Q;Liu J;Tian Y;Liu Y;Wu J
巴巴多斯棉（Gossypium barbadense）是一种棉花栽培品种，具有高纤维品质和抗病原菌（尤其是对主要栽培品种棉的破坏性病原菌Verticilliumdahliae）的抗性。这些优良性状很难通过传统育种方法引入到毛毛草中。此外，G。巴巴多斯尚未成功执行。因此，建立评价G基因功能和分子机制的方法具有重要意义。取得。在本研究中，我们成功地将病毒诱导的基因沉默（VIGS）系统引入了三个葡萄品种。将标记基因插入烟草响尾蛇病毒（TRV）载体。通过优化光强、光周期、苗龄和农杆菌菌株等条件，GaPDS沉默载体侵染的植株100%呈现白色叶片。另外三个标记基因GaCLA1、GaANS和GaANR在G中进一步测试了该VIGS系统。取得。沉默植株的内源基因转录量比对照植株降低了99%以上；这些植株在接种后2周出现表型症状。我们将GaPDS和GaANR基因沉默载体的融合序列片段导入到一株植物中，得到了光漂白和褐色的结果。融合双基因沉默载体浸润植物的沉默程度与单基因沉默载体浸润植物的沉默程度一致。该VIGS系统的开发将促进G基因功能的分析。巴巴多斯，并有助于提供理想的性状育种G。barbadense和G. hirsutum。
Gossypium barbadense is a cultivated cotton species and possesses many desirable traits, including high fiber quality and resistance to pathogens, especially Verticilliumdahliae (a devastating pathogen of Gossypium hirsutum, the main cultivated species). These elite traits are difficult to be introduced into G. hirsutum through classical breeding methods. In addition, genetic transformation of G . barbadense has not been successfully performed. It is therefore important to develop methods for evaluating the function and molecular mechanism of genes in G . barbadense . In this study, we had successfully introduced a virus-induced gene silencing (VIGS) system into three cultivars of G . barbadense by inserting marker genes into the tobacco rattle virus (TRV) vector. After we optimized the VIGS conditions, including light intensity, photoperiod, seedling age and Agrobacterium strain, 100% of plants agroinfiltrated with the GaPDS silencing vector showed white colored leaves. Three other marker genes, GaCLA1, GaANS and GaANR, were employed to further test this VIGS system in G . barbadense . The transcript levels of the endogenous genes in the silenced plants were reduced by more than 99% compared to control plants; these plants presented phenotypic symptoms 2 weeks after inoculation. We introduced a fusing sequence fragment of GaPDS and GaANR gene silencing vectors into a single plant, which resulted in both photobleaching and brownish coloration. The extent of silencing in plants agroinfiltrated with fusing two-gene-silencing vector was consistent with plants harboring a single gene silencing vector. The development of this VIGS system should promote analysis of gene function in G . barbadense , and help to contribute desirable traits for breeding of G . barbadense and G. hirsutum.
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影响因子：
5.1
作者：
Liu, Y;Nakayama, N;Dinesh-Kumar, SP
通讯作者：
Dinesh-Kumar, SP
影响因子：
7.2
作者：
Ratcliff, F;Martin-Hernandez, AM;Baulcombe, DC
通讯作者：
Baulcombe, DC
影响因子：
1.2
作者：
Gao, Xiquan;Britt, Robert C.;He, Ping
通讯作者：
He, Ping
影响因子：
7.2
作者：
Brigneti, G;Martín-Hernández, AM;Jones, JDG
通讯作者：
Jones, JDG
影响因子：
64.5
作者：
Liu, Y;Schiff, M;Dinesh-Kumar, SP
通讯作者：
Dinesh-Kumar, SP