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2025年8月3日 星期日
农业生物技术学报  2024, Vol. 32 Issue (4): 949-960    DOI: 10.3969/j.issn.1674-7968.2024.04.019
  研究资源与技术改进 本期目录 | 过刊浏览 | 高级检索 |
基于PDS基因家族的海马齿VIGS体系的构建
马伟1,3,*, 王熙豪1,*, 张雪妍1,2**
1 海南师范大学 生命科学学院/海南省热带动植物生态学重点实验室,海口571158;
2 海南省东寨港红树林湿地生态系统野外科学观测研究站,海口 570105;
3 海南省海口市长流中学,海口 570312
Construction of Sesuvium portulacastrum VIGS System Based on PDS Gene Family
MA-Wei1,3,*, WANG Xi-Hao1,*, ZHANG Xue-Yan1,2,**
1 College of Life Sciences/Ministry of Education Key Laboratory for Ecology of Tropical Islands, Hainan Normal University, Haikou 571158, China;
2 Hainan Dongzhaigang, Mangrove Ecosystem, Provincial Observation and Research Station, Haikou 570105, China;
3 Hainan Haikou Changliu Middle School, Haikou 570312, China
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摘要 海马齿(Sesuvium portulacastrum)是一种肉质盐生植物,对高盐胁迫具有极强的耐受性,病毒诱导的基因沉默(virus-induced gene silencing, VIGS)是鉴定植物基因功能的重要研究手段,为在耐盐植物海马齿中建立高效的VIGS体系,验证植物耐受逆境胁迫基因的功能,本研究以三亚海马齿为材料,选取八氢番茄脱氢酶(phytoene desaturase, PDS)基因作为标记基因。根据转录组数据通过本地Blast等生物信息学手段鉴定得到5个海马齿PDS家族成员,利用PCR获得5个成员的公共序列(长度为431 bp)构建SpTRV2-PDS沉默表达载体,经根癌农杆菌(Agrobacterium tumefaciens) GV3101转化后通过注射对植株进行侵染,对其表型进行观察,并测定其类胡萝卜素含量变化和PDS基因表达量。结果显示,经注射侵染后,海马齿植株出现生长缓慢及叶片逐渐发黄的症状,7 d后叶片出现白化现象,并可持续存在该表型;与对照组相比,实验组中类胡萝卜素含量和PDS的相对表达量降低,说明成功构建了海马齿的VIGS体系,表明VIGS体系能够有效沉默海马齿中PDS基因。本研究为鉴定海马齿耐盐基因功能提供了技术手段。
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马伟
王熙豪
张雪妍
关键词 海马齿八氢番茄脱氢酶(PDS)基因家族序列分析病毒诱导的基因沉默(VIGS)载体构建    
AbstractSesuvium portulacastrum is a succulent saline plant, which is extremely tolerant to high salt stress. Virus-induced gene silencing (VIGS) is an important research tool to characterise the gene function of plants. In order to establish an efficient VIGS system in the important salt-tolerant plant S. portulacastrum, phytoene desaturase (PDS) gene was selected as a marker gene using S. portulacastrum of Sanya as the material. According to the transcriptome data, 5 PDS family members of S. portulacastrum were identified by local Blast and other biosignature means, and the common sequences of the 5 members were obtained by PCR, with a length of 431 bp.Meanwhile, SpTRV2-PDS silencing expression vector was constructed, which was transformed by Agrobacterium tumefaciens GV3101 and then infiltrated into the plants by injection. The phenotype was observed, and the changes of carotenoid content and PDS gene expression were measured. The results showed that after infection, the plants showed slow growth and gradual yellowing of leaves, and the leaves showed albino phenomenon after 7 d, and the carotenoid content and the relative expression of PDS in the experimental group continued to exist, indicating that the VIGS system of S. portulacastrum was successfully constructed, indicating that the VIGS system could effectively silence the expression of PDS gene in S. portulacastrum. This study provides a technical means for the identification of the function of salt tolerance genes in S. portulacastrum.
Key wordsSesuvium portulacastrum    Phytoene desaturase (PDS) gene family    Sequence analysis    Virus-induced gene silencing (VIGS)    Vector construction
收稿日期: 2023-06-12     
ZTFLH:  S3  
基金资助:海南省自然科学基金(2019RC185)
通讯作者: **zhangxueyan_caas@126.com   
作者简介: *同等贡献作者
引用本文:   
马伟, 王熙豪, 张雪妍. 基于PDS基因家族的海马齿VIGS体系的构建[J]. 农业生物技术学报, 2024, 32(4): 949-960.
MA-Wei, WANG Xi-Hao, ZHANG Xue-Yan. Construction of Sesuvium portulacastrum VIGS System Based on PDS Gene Family. 农业生物技术学报, 2024, 32(4): 949-960.
链接本文:  
https://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2024.04.019     或     https://journal05.magtech.org.cn/Jwk_ny/CN/Y2024/V32/I4/949
 
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