温度和土壤水分与Bt棉铃壳中杀虫蛋白表达关系及其氮代谢生理机制

doi:10.3864/j.issn.0578-1752.2018.07.004

摘要/Abstract

摘要： 【目的】明确温度和土壤水分对Bt棉杀虫蛋白含量及其氮代谢活性的影响，为生产中Bt棉抗虫性的安全稳定利用提供理论参考。【方法】2016—2017年以转Bt抗虫基因抗虫棉常规品种泗抗1号（SK1）和杂交种泗抗3号（SK3）为材料，采用盆栽法，设置29℃、32℃、35℃、38℃ 4个温度水平，土壤最大持水量的80%、70%、60%、50%、40% 5个土壤水分水平，观察温度和土壤水分对Bt棉铃壳杀虫蛋白含量的影响，各处理持续胁迫4 d。2016年主要研究各处理对Bt棉铃壳中杀虫蛋白含量的影响；在此基础上，2017年进一步探讨各处理对铃壳中可溶性蛋白含量、谷氨酸草酰乙酸转氨酶（GOT）、蛋白酶、肽酶活性的影响。【结果】SK1和SK3杀虫蛋白含量均在32℃、最大持水量为60%时最高，分别达到471.1 ng·g^-1 FW和351.7 ng·g^-1 FW。在同一土壤水分条件下，32℃最利于SK1和SK3杀虫蛋白表达；同一温度条件下，最大持水量60%利于SK1和SK3杀虫蛋白表达。对杀虫蛋白含量与温度和土壤水分关系进行二元多项式回归分析发现，Bt棉杀虫蛋白含量（Y）与温度（X₂）和土壤水分（X₁）呈二元二次方程关系，其中SK1、SK3相关方程分别为Y=-3230.2+17.2X₁+199.1X₂-0.3X₁²-3.7X₂²-0.7X₁X₂（r＝0.829^**）、Y=-3322.0+40.7X₁+145.2X₂-0.3X₁²-2.0X₂²-0.3X₁X₂（r＝0.739^**）。SK1的杀虫蛋白表达量最大的温度和土壤水分条件为31.8℃、57.8%，SK3为33.2℃、60.8%。氮代谢相关生理特征表明，SK1和SK3均表现为在32℃和土壤含水量为60%处理下，棉铃中可溶性蛋白含量、谷氨酸草酰乙酸转氨酶（GOT）活性较高，蛋白酶、肽酶活性较低；杀虫蛋白含量与可溶性蛋白和GOT活性呈极显著正相关关系（r＝0.613**；r＝0.735**），与蛋白酶活性和肽酶活性呈极显著负相关关系（r＝-0.724**；r＝-0.738**）。【结论】温度和土壤水分通过调控蛋白质分解和合成，共同影响Bt棉杀虫蛋白表达，且与其含量呈二元二次方程关系。

关键词: Bt棉, 温度, 土壤水分, 杀虫蛋白, 氮代谢

Abstract: 【Objective】 The effects of different temperature and soil moisture on the insecticidal protein contents in Bacillus thuringiensis (Bt) cotton boll shell were investigated to provide a theoretical reference for the safe and stable utilization of Bt cotton in production.【Method】The study was undertaken on two Bt cotton cultivars Sikang 1 (SK1, a conventional cultivar) and Sikang 3 (SK3, a hybrid cultivar) during 2016 and 2017 growing seasons at Yangzhou University Farm, Yangzhou, China. The study was arranged with two factors that consisted of four temperature (29, 32, 35, and 38℃) and five soil moisture content (40%, 50%, 60%, 70% and 80% field capacity). The potted cotton plants were exposed to the twenty treatments for 4 days. In 2016, the effects of temperature and soil moisture on the insecticidal protein contents in boll shell were determined, and the soluble protein contents, glutamate oxaloacetate transaminase (GOT) activities, protease and peptidase activities were further studied in 2017. 【Result】The highest contents of insecticidal protein for SK1 and SK3 were both observed under the treatment of 32℃/60% field capacity, which was 471.1 ng·g^-1 FW and 351.7 ng·g^-1 FW, respectively. Under the same soil moisture, the insecticidal protein contents for SK1 and SK3 at 32℃ were significantly higher than those for other three temperature conditions. Under the same temperature, the insecticidal protein content under 60% field capacity for SK1 and SK3 was higher. Polynomial regression analysis showed a quadratic relationship between the insecticidal protein contents and the temperature and soil moisture. The binary quadric fitting equations for SK1 and SK3 were: Y=-3230.2+17.2X₁+199.1X₂-0.3X₁²-3.7X₂²-0.7X₁X₂(r＝0.829^**), Y=-3322.0+40.7X₁+145.2X₂- 0.3X₁²-2.0X₂²-0.3X₁X₂(r＝0.739^**), respectively, where Y stands for insecticidal protein contents, and X₁ is soil moisture, X₂is temperature. Based on the equations, the maximum insecticidal protein contents would be obtained under the combination of 31.8℃/57.8% for SK1 and 33.2℃/60.8% for SK3, respectively. The physiological characteristics of nitrogen metabolism showed that the soluble protein contents and the GOT activities were highest in the boll shell for SK1 and SK3 under the combination of 32℃/60% field capacity, whereas the lowest protease and peptidase activities were also detected under this condition. The correlation analysis showed that there was a significant positive correlation between the insecticidal protein contents with soluble protein content and GOT activity (r= 0.613**; r= 0.735**), while the insecticidal protein was negatively correlated with protease activity and peptidase activity (r= -0.724**; r= -0.738**).【Conclusion】 The temperature and soil moisture affected the expression of Bt insecticidal protein by regulating the protein synthesis and degradation. And the relationship between Bt protein contents and temperature and soil moisture could be quantified by a binary quadric fitting equation.

Key words: Bt cotton, temperature, soil moisture, insecticidal protein, nitrogen metabolism

张祥，王剑，彭盛，芮秋治，李丽楠，陈媛，陈源，陈德华. 温度和土壤水分与Bt棉铃壳中杀虫蛋白表达关系及其氮代谢生理机制[J]. 中国农业科学, 2018, 51(7): 1261-1271.

ZHANG Xiang, WANG Jian, PENG Sheng, RUI QiuZhi, LI LiNan, CHEN Yuan, CHEN Yuan, CHEN DeHua. Relationship Between Temperature, Soil Moisture, and Insecticidal Protein Content in Bt Cotton Boll Shell and the Mechanism of Nitrogen Metabolism[J]. Scientia Agricultura Sinica, 2018, 51(7): 1261-1271.

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