Jaburetox 2-Ec 在马铃薯中的表达编码了对 Tuta absoluta Meyrick(鳞翅目:蝼蛄)的抗性

IF 2.3 3区 农林科学 Q1 AGRONOMY Potato Research Pub Date : 2024-09-14 DOI:10.1007/s11540-024-09788-z
Merve Tekinsoy, Allah Bakhsh, Mehmet Emin Çalışkan
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引用次数: 0

摘要

番茄潜叶蝇 Tuta absoluta Meyrick 是全世界最严重的马铃薯害虫。在本研究中,我们探索了另一种诱导马铃薯对 T. absoluta 产生抗性的方法,即通过工程化 Jaburetox-2Ec 来诱导马铃薯对 T. absoluta 产生抗性,Jaburetox-2Ec 是一种杀虫肽,来源于杰克豆(Canavalia ensiformis L.)脲酶的一种同工酶。该构建体是在 35S 启动子控制下,将 Jaburetox-2Ec 的 PCR 扩增片段(270 bp)克隆到 pBIN61 载体中,并命名为 pBIN-JBTX。此外,将表达 Jaburetox-2Ec 的重组质粒转化到马铃薯变种 Lady Olympia 中。此外,还利用农杆菌菌株 EHA105 将表达 Jaburetox-2Ec 的重组质粒转化到马铃薯品种 Lady Olympia 中。结果表明,T-DNA 区域在初代转化子中正确整合。定量 PCR 检测表明,独立转基因品系的 Jaburetox 转录水平有所提高。叶片生物毒性试验是通过在转基因马铃薯植株上释放预先饥饿的绝对蓟马二龄、三龄和四龄幼虫进行的。结果发现,根据幼虫阶段的不同,幼虫取食转基因植物的死亡率在 11% 到 33% 之间。此外,研究还发现,取食对照植株的幼虫比取食转基因植株的幼虫体重增加得更多。此外,还记录到取食转基因植物的幼虫出现畸形。总之,研究结果表明 Jaburetox 2-Ec 对绝对拟南芥具有杀虫活性。转基因品系可作为高效马铃薯育种计划的种质资源。
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Expression of Jaburetox 2-Ec in Potato Encodes Resistance Against Tuta absoluta Meyrick (Lepidoptera: Gelechiidae)

The tomato leaf miner Tuta absoluta Meyrick is the most serious pest of potato worldwide. In the present study, we explored an alternate approach to induce resistance in potatoes against T. absoluta by engineering Jaburetox-2Ec, an insecticidal peptide derived from an isoform of urease from Jack bean (Canavalia ensiformis L.). The construct was developed by cloning a PCR-amplified fragment of Jaburetox-2Ec (270 bp) in a pBIN61 vector under the control of a 35S promoter and was named pBIN-JBTX. Furthermore, the recombinant plasmid expressing Jaburetox-2Ec was transformed into potato cv. Lady Olympia using Agrobacterium tumefaciens strain EHA105. The results showed the proper integration of the T-DNA region in primary transformants. Quantitative PCR assays showed the independent transgenic lines had enhanced transcript levels of Jaburetox. The leaf biotoxicity assays were performed by releasing pre-starved 2nd, 3rd, and 4th instar larvae of T. absoluta on transgenic potato plants. As a result, depending on the larval stage, the mortality rate of larvae feeding on transgenic plants ranged from 11 to 33%. Additionally, it was found that larvae feeding on control plants gained more weight than those feeding on transgenic plants. Besides that, deformities in larvae feeding on transgenic plants were also recorded. Overall, the results exhibited the insecticidal activity of Jaburetox 2-Ec against T. absoluta. The transgenic lines can be used as a source of germplasm for an efficient potato breeding programme.

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来源期刊
Potato Research
Potato Research AGRONOMY-
CiteScore
5.50
自引率
6.90%
发文量
66
审稿时长
>12 weeks
期刊介绍: Potato Research, the journal of the European Association for Potato Research (EAPR), promotes the exchange of information on all aspects of this fast-evolving global industry. It offers the latest developments in innovative research to scientists active in potato research. The journal includes authoritative coverage of new scientific developments, publishing original research and review papers on such topics as: Molecular sciences; Breeding; Physiology; Pathology; Nematology; Virology; Agronomy; Engineering and Utilization.
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