Conversion of sheath blight susceptible indica and japonica rice cultivars into moderately resistant through expression of antifungal β-1,3-glucanase transgene from Trichoderma spp.

IF 2.7 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Transgenic Research Pub Date : 2022-10-01 Epub Date: 2022-08-09 DOI:10.1007/s11248-022-00318-6
Shivali Pathania, Jagjeet Singh Lore, Anu Kalia, Ajinder Kaur, Manveer Sharma, Gurjit Singh Mangat, Jagdeep Singh Sandhu
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引用次数: 2

Abstract

Rice is an important food crop for three billion people worldwide. The crop is vulnerable to several diseases. Sheath blight caused by fungal pathogen Rhizoctonia solani is a significant threat to rice cultivation accounting for up to 50% yield losses. The pathogen penetrates leaf blades and sheaths, leading to plant necrosis; and major disease resistance gene against the pathogen is not available. This study describes development of sheath blight resistant transgenic indica and japonica rice cultivars through introduction of antifungal β-1,3-glucanase transgene cloned from Trichoderma. The transgene integration and expression in transformed T0 rice plants was examined by PCR, RT-PCR, qRT-PCR demonstrating up to 5-fold higher expression as compared to non-transgenic plants. The bioassay of T0, T1 and homozygous T2 progeny plants with virulent R. solani isolate revealed that plants carrying high level of β-1,3-glucanase expression displayed moderately resistant reaction to the pathogen. The optical micrographs of leaf sheath cells from moderately resistant plant after pathogen inoculation displayed presence of a few hyphae with sparse branching; on the contrary, pathogen hyphae in susceptible non-transgenic plant cells were present in abundance with profuse hyphal branching and forming prominent infection cushions. The disease severity in T2 progeny plants was significantly less as compared to non-transgenic plants confirming role of β-1,3-glucanase in imparting resistance.

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通过表达木霉抗真菌β-1,3-葡聚糖酶基因将易受纹枯病影响的籼稻和粳稻品种转化为中等抗性品种。
水稻是全球30亿人口的重要粮食作物。这种作物易受几种病害的侵害。由真菌病原菌索拉丝核菌引起的纹枯病是水稻栽培的重大威胁,可造成高达50%的产量损失。病原菌渗入叶片和鞘,导致植物坏死;目前还没有针对病原菌的主要抗病基因。本研究通过引入从木霉中克隆的抗真菌β-1,3-葡聚糖酶基因,来开发抗纹枯病转基因籼稻和粳稻品种。通过PCR、RT-PCR和qRT-PCR检测了转基因水稻植株的整合和表达,结果显示转基因水稻植株的表达量比非转基因植株高5倍。对毒力极强的茄茄疫病毒株的T0、T1和T2纯合子植株进行生物测定,结果表明,高表达β-1,3-葡聚糖酶的植株对该病原菌表现出中度抗性反应。中等抗性植物接种病原菌后叶鞘细胞光学显微镜显示菌丝较少,分枝稀疏;而在非转基因易感植物细胞中,病原菌菌丝大量存在,菌丝分支丰富,形成明显的感染垫。与非转基因植株相比,T2后代植株的疾病严重程度显著降低,证实了β-1,3-葡聚糖酶在传递抗性中的作用。
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来源期刊
Transgenic Research
Transgenic Research 生物-生化研究方法
CiteScore
5.40
自引率
0.00%
发文量
38
审稿时长
4-8 weeks
期刊介绍: Transgenic Research focusses on transgenic and genome edited higher organisms. Manuscripts emphasizing biotechnological applications are strongly encouraged. Intellectual property, ethical issues, societal impact and regulatory aspects also fall within the scope of the journal. Transgenic Research aims to bridge the gap between fundamental and applied science in molecular biology and biotechnology for the plant and animal academic and associated industry communities. Transgenic Research publishes -Original Papers -Reviews: Should critically summarize the current state-of-the-art of the subject in a dispassionate way. Authors are requested to contact a Board Member before submission. Reviews should not be descriptive; rather they should present the most up-to-date information on the subject in a dispassionate and critical way. Perspective Reviews which can address new or controversial aspects are encouraged. -Brief Communications: Should report significant developments in methodology and experimental transgenic higher organisms
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