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20222023

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1Expression and synergistic activity of enhancin from Pseudaletia unipuncta granulovirus-Ps in Bacillus thuringiensis

HUANG Lixin,SUN Jun,LIU Qin, HAN Guangjie,LI Chuanming,XIA Yang,LU Yurong,XU Jian

[Objective] To express the truncated fragments of enhancin gene from Pseudaletia unipuncta granulovirus-Ps（PuGV-Ps） in Bacillus thuringiensis（Bt） and provide a theoretical basis for the construction of Bt engineering bacteria. [Methods] The codon of the truncated fragments of enhancin gene was optimized for the construction of the expression vectors of enhancin and the expression of fusion proteins. Then, the expression levels of enhancin under the guidance of two promoters were analyzed, and the synergistic activity of enhancin on Bt was determined. [Results] The expression vectors pHTP cry1Ac CoEn81, pHTRHCoEn81, and pHTNCCoEn81 were constructed in the study. SDS-PAGE showed that pHTP cry1Ac CoEn81 and pHTNCCoEn81 produced recombinant proteins of 81 kDa and 134 kDa, respectively. There was no significant difference in the expression level of enhancin or the yield of recombinant protein under the guidance of promoters P cry1Ac and Pcry8E. Furthermore, the recombinant enhancin significantly increased the insecticidal activity of Bt against Plutella xylostella.[Conclusion] The codon-optimized enhancin of PuGV-Ps can be expressed in Bt and has significant synergistic activity, which provides theoretical guidance for the construction and application of Bt engineering bacteria with high efficiency.

2023Acta Microbiologica Sinica（2023）

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2Effects of Piriformospora indica on the Growth and Antioxidant System of Rice Seedlings Under Salt Stress

XIA Yang,LI Chuanming,LIU Qin, HAN Guangjie, XU Bin,HUANG Lixin,QI Jianhang,LU Yurong,XU Jian

[目的]探究印度梨形孢(Piriformospora indica)PI-020对盐胁迫下水稻幼苗生长、抗氧化相关酶活性及基因表达水平的影响.[方法]以不同浓度的PI-020菌丝体悬液接种南粳9108水培苗,显微镜观察及qPCR技术分析PI-020在水稻秧苗根系的定殖能力,并测定水稻叶片丙二醛(MDA)含量,分析水稻秧苗的表型参数、光合色素含量、抗氧化相关酶活性及基因表达的变化.[结果]与对照相比,盐胁迫条件下(100 mmol/L NaCl),PI-020定殖后,水稻叶片MDA含量显著降低.200倍菌丝体稀释液处理的效果最好,MDA含量减少了67.2％.PI-020定殖后,水稻株高、根长、叶面积、鲜质量和干质量分别增加了34.67％、23.62％、58.04％、59.53％和67.25％,与对照相比均差异显著;同时PI-020定殖还显著提高了叶片光合色素含量,抗氧化酶CAT、APX、POD活性及抗氧化相关基因OsCAT、OsAPX2、OsSOD的表达水平.[结论]印度梨形孢PI-020通过提高水稻幼苗抗氧化能力减少盐胁迫引起的氧化损伤,从而降低MDA含量,同时缓解光合色素的降解,保护了水稻光合系统,进而提高水稻耐盐性.

2023Chinese Journal of Rice Science（2023）引用：1

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3Sub-lethal Effect of Cnaphalocrocis Medinalis Granulovirus to the Growth, Reproduction and Physiology of Host

LIU Qin,LI Chuanming, HAN Guangjie,XU Bin,HUANG Lixin,LU Yurong,XIA Yang,XU Jian

稻纵卷叶螟颗粒体病毒(CnmeGV)是感染稻纵卷叶螟的专性杆状病毒.明确CnmeGV亚致死作用对稻纵卷叶螟生长发育、繁殖能力和生理生化的影响,为应用CnmeGV防治稻纵卷叶螟提供依据.CnmeGV 1×105 OB/g浓度处理3龄稻纵卷叶螟幼虫10 d后存活幼虫虫体内仍普遍检测到CnmeGV的存在,从而产生亚致死作用.亚致死CnmeGV处理对稻纵卷叶螟的生长发育产生明显影响,主要表现在变态发育的化蛹、羽化和产卵过程,存活个体蛹重减轻了15.95％,羽化率下降了24.81％,产卵量减少41.5％,差异显著.同时雌雄性比从1.11下降到0.82.CnmeGV亚致死处理诱导幼虫能量代谢增强,中肠消化酶中淀粉酶、脂肪酶活性显著提高,而解毒酶中羧酸酯酶活性显著下降,谷胱甘肽转移酶活性显著提高.CnmeGV亚致死作用影响到虫体激素分泌,幼虫体内蜕皮激素、保幼激素水平较对照分别下降了4.59和5.62倍.CnmeGV亚致死处理雌成虫卵黄蛋白含量下降了40.63％,可能是导致产卵量降低的主要因素.

2022Chinese Journal of Biological Control（2022）

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4Isolation and Identification of Entomopathogenic Nematode N-Yz1 from Chilo Suppressalis and Its Infection Characteristics

LIU Qin,XIA Yang, HAN Guangjie,LI Chuanming,LU Yurong,HUANG Lixin,QI Jianhang,XU Jian

[目的]二化螟是危害水稻的重要害虫,其钻蛀为害,防治困难,严重威胁水稻生产.本研究对感染二化螟昆虫病原线虫的分离筛选和感染特性进行研究,期望为水稻钻蛀害虫危害的防治提供新的途径和方法.[方法]自越冬代二化螟幼虫体内通过组织分离筛选,获得野外感染二化螟的昆虫病原线虫,室内接种实验分析分离昆虫病原线虫对二化螟的感染活性和作用特性.[结果]自然罹死二化螟幼虫体内分离获得野外感染的昆虫病原线虫N-Yz1,接种4龄二化螟幼虫导致感染并死亡,后期虫体内产生大量形态一致的侵染期线虫(IJ).形态学观察结合rDNA-ITS序列分析,鉴定分离昆虫病原线虫隶属于斯氏线虫属,与已鉴定种小卷蛾斯氏线虫同源性达99.87％.昆虫病原线虫N-Yz1对不同龄期二化螟幼虫都有较高感染致死作用,40头(IJ)的线虫接种剂量,对低龄(2～3龄)二化螟致死速度快、致死率高,接种24 h就出现感染致死个体,48 h后幼虫全部死亡.高龄幼虫(4～6龄)感染致死速度较慢,死亡高峰发生在接种线虫后的48 h.N-Yz1接种剂量与二化螟死亡率呈显著正相关,剂量超过30 IJs/头时,4龄二化螟感染死亡率达90％以上.N-Yz1对二化螟幼虫具有明显搜索能力,在距离寄主害虫0-30 cm的范围内,通过迁移扩散,感染二化螟并导致其死亡.[结论]分离昆虫病原线虫N-Yz1具有较强的感染致死能力,表现出防治水稻钻蛀害虫二化螟的潜在应用价值.

2022

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