VIGS技术鉴定木薯糖转运蛋白Mesweet18的功能研究

doi:10.19586/j.2095-2341.2022.0075

生物技术进展 ›› 2022, Vol. 12 ›› Issue (6): 888-893.DOI: 10.19586/j.2095-2341.2022.0075

VIGS技术鉴定木薯糖转运蛋白Mesweet18的功能研究

薛晶晶(), 安飞飞, 罗秀芹, 蔡杰()

中国热带农业科学院热带作物品种资源研究所，农业农村部木薯种质资源保护与利用重点实验室，海口 571131

收稿日期:2022-05-13 接受日期:2022-07-12 出版日期:2022-11-25 发布日期:2022-11-30
通讯作者: 蔡杰
作者简介:薛晶晶E-mail: xuetao608@163.com；
基金资助:
海南省基础与应用基础研究计划（自然科学领域）高层次人才项目(2019RC314)

Study on the Function of Cassava Sugar Transporter Mesweet18 by VIGS

Jingjing XUE(), Feifei AN, Xiuqin LUO, Jie CAI()

Key Laboratory of Conservation and Utilization for Cassava Germplasm Resources，Tropical Crops Genetic Resources Institute，Chinese Academy of Tropical Agricultural Sciences，Haikou 571131，China

Received:2022-05-13 Accepted:2022-07-12 Online:2022-11-25 Published:2022-11-30
Contact: Jie CAI

摘要/Abstract

摘要：

糖转运蛋白（sugar will eventually be exported transporter，SWEET）在植物运输糖类、生殖和发育、逆境性、与病原体互作等方面发挥着重要作用。选择木薯糖转运蛋白Mesweet18基因沉默的靶基因区域，通过病毒诱导的基因沉默(virus-induced gene silencing，VIGS)技术注射木薯SC9的盆栽苗叶片。qRT-PCR结果表明，Mesweet18在沉默植株中的表达量显著下调，分别是对照的46.80%、30.23%、21.12%。叶片叶绿素和可溶性糖含量检测结果表明，与对照相比，叶绿素a、b和总含量均出现不同程度的下降，蔗糖和果糖含量显著增加，而葡萄糖含量出现轻微下降。研究Mesweet18在木薯中的分子功能，为深入研究糖转运蛋白SWEET在木薯中的分子机制奠定了基础。

关键词: 木薯, SWEET转运蛋白, VIGS, 可溶性糖含量

Abstract:

Sugar will eventually be exported transporter （SWEET） is a key protein for plants， which plays an important role in plant sugar transport， reproduction and developnent， stress resistance， and interaction with pathogens. In this study， we used potted plantings of cassava SC9 as materials， designed the target gene region of Mesweet18 silencing in cassava， used virus-induced gene silencing （VIGS） technology to infect. qRT-PCR results showed that the expression of Mesweet18 in silencing plant were 46.80%，30.23%，21.12%， respectively， which were significantly down-regulated than the control. The results of chlorophyll and soluble sugar content in leaves showed that chlorophyll a， b and total content were decreased， sucrose and fructose contents increased significantly， while glucose content decreased slightly， which compared with the control. The study on molecular function of Mesweet18 in cassava laid a foundation for the molecular mechanism of sugar transporter SWEET in cassava.

Key words: cassava, SWEET transporter, VIGS, soluble sugar content

中图分类号:

Q533

薛晶晶, 安飞飞, 罗秀芹, 蔡杰. VIGS技术鉴定木薯糖转运蛋白Mesweet18的功能研究[J]. 生物技术进展, 2022, 12(6): 888-893.

Jingjing XUE, Feifei AN, Xiuqin LUO, Jie CAI. Study on the Function of Cassava Sugar Transporter Mesweet18 by VIGS[J]. Current Biotechnology, 2022, 12(6): 888-893.

图/表 5

表1 研究所用引物序列

Table 1 Primer sequences used in the study

引物名称	引物序列(5'→3')
VIGS-Mesweet18-F	5'-AGTGGTCTCTGTCCAGTCCTTGGAACATTTTGGAGAAT-3'
VIGS-Mesweet18-R	5'-GGTCTCAGCAGACCACAAGTACTTGACCCTGCAATGCT-3'
qPCR-Mesweet18-F	5'-GGAGTTTGAGAGTCTTCCTTATGTG-3'
qPCR-Mesweet18-R	5'-GAGTTACATAGACGATTTCCACCAG-3'
qPCR-Actin-F	5'-TGATGAGTCTGGTCCATCCA-3'
qPCR-Actin-R	5'-CCTCCTACGACCCAATCTCA-3'
CsCMV-F	5'-TGGGCGCTAATTAGTTTACTGCA-3'
CsCMV-R	5'-GGTCAAGACGGCTCAACTCTTCA-3'

图1 木薯Mesweet18 沉默植株电泳检测注: M-DNA Marker；1—空白对照WT；2—阴性对照pCsCMV-NC；3~5—实验组；泳道6~7—阳性对照pCsCMV-CHLD。

Fig. 1 Electrophoresis detection of cassava Mesweet18 silenced plants

图2 木薯Mesweet18沉默植株表型观察

Fig. 2 Phenotypic observation of cassava Mesweet18 silenced plants

图3 木薯Mesweet18 沉默植株的表达水平分析注：**表示与空白对照组相比在P<0.01水平上具有统计学意义。

Fig. 3 Expression level analysis of cassava Mesweet18 silenced plants

图4 木薯Mesweet18沉默植株叶片中叶绿素和可溶性糖含量分析注：*和**分别表示与空白对照组相比不同处理在P<0.05和P<0.01水平上具有统计学意义。

Fig. 4 Analysis of chlorophyll and soluble sugar content in cassava Mesweet18 silenced plants

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VIGS技术鉴定木薯糖转运蛋白Mesweet18的功能研究

Study on the Function of Cassava Sugar Transporter Mesweet18 by VIGS

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