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【目的】研究土壤水分亏缺对Bt棉杀虫蛋白含量及Bt基因表达、氮代谢酶活性的影响,为Bt棉抗虫性安全表达提供理论参考。【方法】2014—2015年以Bt棉常规品种泗抗1号、杂交种泗抗3号为材料,采用盆栽法,2014年设置5个土壤水分处理:G1、G2、G3、G4和CK,其土壤含水量分别为最大持水量的15%、30%、45%和75%。2015年设置4个处理:G2、G3、G4和CK。观察土壤水分亏缺对盛铃期Bt棉铃壳杀虫蛋白含量影响。所有处理于盛花期前10 d控制浇水,如遇下雨,将处理盆钵移入室内。使用WET土壤三参数速测仪监测土壤水分,用称重法控制土壤水分,即当监测发现土壤水分低于设计值时,于早晨、中午、傍晚进行定量补水。2015年进一步研究水分亏缺对Bt基因表达量、氮代谢相关合成酶(硝酸还原酶和谷氨酸丙酮酸转氨酶)活性、分解酶(肽酶和蛋白酶)活性的影响。【结果】与对照(土壤含水量为最大持水量75%)相比,泗抗1号和泗抗3号铃壳中Bt蛋白含量随土壤水分亏缺程度的增加而降低,且在土壤含水量为最大持水量60%时开始显著下降,但泗抗1号下降幅度低于泗抗3号,其中2014年泗抗1号下降22.5%,泗抗3号下降41.6%。在土壤含水量为最大持水量60%时,铃壳中Bt基因表达量增加,泗抗1号、泗抗3号分别比对照提高48.6%和22.1%。氮代谢相关酶活性变化表明,水分亏缺条件下,2个类型品种的硝酸还原酶(NR)和谷氨酸丙酮酸转氨酶(GPT)活性降低,肽酶和蛋白酶活性增加。且肽酶和蛋白酶活性变化幅度高于NR和GPT。相关分析表明,NR和GPT活性与铃壳中杀虫蛋白含量呈显著或极显著正相关;肽酶和蛋白酶活性与杀虫蛋白含量呈显著负相关。【结论】水分亏缺胁迫下,供试品种铃壳中杀虫蛋白质含量下降。但在转录水平,未发现Bt基因表达量下降。但氮代谢关键合成酶(NR和GPT)活性降低,分解酶(肽酶和蛋白酶)活性增加。因此,蛋白质合成减弱、分解加强可能导致铃壳中杀虫蛋白含量下降。 相似文献
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为评价外源Bt基因插入以及氮肥对Bt棉功能叶Bt蛋白表达及氮代谢的影响,对Bt棉功能叶Bt蛋白含量,硝酸还原酶、谷丙转氨酶和蛋白酶活性,可溶性蛋白质和全氮含量等指标进行了测定。结果表明:Bt棉功能叶Bt蛋白含量在蕾期最高,此后,随生育进程呈显著下降。与常规棉相比,Bt棉功能叶硝酸还原酶活性提高,在前中期谷丙转氨酶活性显著提高且有利于可溶性蛋白质的合成,前期全氮含量显著增加,对蛋白酶活性没有明显影响。Bt棉功能叶Bt蛋白含量与硝酸还原酶活性、可溶性蛋白质和全氮含量呈显著正相关,表明Bt棉前期增强的氮代谢促进了Bt蛋白的表达。施氮肥明显提高Bt棉中后期功能叶硝酸还原酶活性、可溶性蛋白质含量和Bt蛋白的表达。基施和初花期各50%的效应显著大于全部基施,而在基施氮肥基础上,初花期增施50%的氮肥效应最明显。与常规棉比较,施氮肥更有利于Bt棉中后期功能叶硝酸还原酶活性、可溶性蛋白质含量提高。 相似文献
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为了探讨种植密度对转基因抗虫棉纤维中杀虫(Bt)蛋白表达量的影响及其氮代谢机制,以Bt棉常规种泗抗1号和Bt棉杂交种泗抗3号为试验材料,比较1.5×104、3.0×104、4.5×104、6.0×104、7.5×104 株·hm-2五个种植密度下转Bt棉杀虫蛋白表达量及其氮代谢的差异。结果表明,纤维中Bt蛋白表达量随种植密度增大呈上升趋势。相关分析表明,单株铃数、单铃体积、单铃干重与Bt蛋白表达量均呈显著负相关。纤维中氮代谢生理相关指标中,可溶性蛋白含量及谷氨酸丙酮酸转氨酶(GPT)和谷氨酸草酰乙酸转氨酶(GOT)活性随种植密度的增大而升高,但游离氨基酸含量、蛋白酶和肽酶活性随密度增大而下降。纤维中Bt蛋白表达量与可溶性蛋白含量、GPT和GOT活性呈显著正相关,而与游离氨基酸含量、蛋白酶和肽酶活性呈显著负相关。可见,提高种植密度降低了棉铃生长,但提高了纤维中Bt蛋白的表达量,且Bt蛋白表达量的提高与植株氮代谢生理密切相关。因此,在生产中通过协调种植密度既可以保持棉铃适度发育,又有利于提高棉纤维中Bt蛋白的表达量,为提高Bt棉产量器官抗虫性提供栽培基础。 相似文献
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以常规种泗抗1号(SK-1)和杂交种泗抗3号(SK-3)作为试验材料,设置喷清水(CK)和不同浓度(1.0%～10.0%)尿素处理,研究喷施不同浓度尿素对Bt棉蕾中杀虫蛋白表达量的影响。结果表明：随着尿素浓度的增加,棉蕾中杀虫蛋白含量呈先增加后降低的趋势,尿素浓度7.0%～8.0%处理的棉蕾中杀虫蛋白含量最高,较对照增加14.9%～24.8%。氮代谢机制表明,棉蕾中可溶性蛋白质和游离氨基酸含量增加,谷氨酰胺合成酶(GS)、谷氨酸合酶(GOGAT)、谷草转氨酶(GOT)和谷丙转氨酶(GPT)活性提高,与Bt杀虫蛋白含量表现一致;蛋白酶和肽酶活性与Bt蛋白含量量呈相反特征。主成分分析进一步表明,两品种棉蕾的可溶性蛋白质和游离氨基酸含量,GS、GOGAT、GOT和GPT活性对Bt杀虫蛋白含量变化的贡献分别达71.8%和72.4%;蛋白酶和肽酶活性对Bt杀虫蛋白含量变化的贡献分别为23.4%和21.6%。综上,蕾期根外喷施尿素,Bt棉蕾的蛋白质合成提高是杀虫蛋白含量提高的主要原因。 相似文献
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以转Bt基因抗虫棉小铃品种泗抗1号和大铃品种科棉6号为材料,于2009～2010年在扬州大学实验农牧场进行了源库调节对棉铃氮代谢影响的试验。结果表明,与对照相比,去除棉株每张叶片的一半后,体积膨大期棉铃铃重变小,氮代谢有关化合物含量和酶活性提高,且大铃品种科棉3号受影响较大;整株均匀去除一半蕾,则使得铃重变大,氮代谢有关化合物含量和酶活性下降,仍然是大铃品种受影响较大。相关分析结果表明,源库调节下体积增大期棉铃氮代谢强度与铃重基本上呈显著或极显著负相关。因此转Bt基因棉花群体源小库大或源大库小均不利于棉铃生理氮代谢与棉铃发育的协调,同样不利于产量和抗虫性的协同表达。 相似文献
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通过检测分析,转Bt基因抗虫棉金杂棉3号是抗棉铃虫优良的新品种,在棉花不同的生育期,Bt杀虫蛋白含量幅度在94.2-896.8 ng·g^-1,苗期、蕾期、花期的叶片Bt蛋白表达量级属于高表达,蕾期的小蕾和花期的花瓣Bt蛋白表达量级属于中高表达,花期的小蕾、铃期花瓣和小蕾Bt蛋白表达量级属于中表达,花期的小铃、铃期叶片和小铃Bt蛋白表达量级属于低表达。金杂棉3号的杀虫活性80.21%-91.87%,抗虫效率达83.33%-100.00%,保蕾效果达82.56%-100.00%。 相似文献
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通过南疆膜下滴灌棉花不同灌水量和施氮量的田间试验，研究了棉花在不同生育时期及不同叶位叶片叶绿素含量的时空分布。结果表明，在苗期氮肥用量对子叶叶绿素含量的影响显著，随叶位增加，对真叶叶绿素含量的影响越来越小。盛蕾期是棉花叶片叶绿素含量变化最为敏感的时期，并受氮肥施用量的显著影响。在花铃期氮肥用量和叶位均影响棉花叶片叶绿素含量，同时施氮量和滴水量的比值对叶绿素含量的影响也极为明显；在盛铃期，水分与氮肥施用量对棉花叶绿素含量的影响已不显著，但不同叶位之间仍然差异很大。在棉花生长的中后期，滴水量主要影响棉花下部叶片叶绿素含量，而氮素供应量主要影响上部叶片叶绿素含量，中部叶片叶绿素含量同时受水分和氮素的调节。南疆膜下滴灌棉花叶片叶绿素含量，在苗期和盛蕾期以下部叶片最高，花铃期和盛铃期以中部叶片最高，每个叶位叶绿素含量均随着生育时期的推移而逐渐增加。 相似文献
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氮素营养水平对黄瓜氮代谢关键酶活性变化及氮化物的影响 总被引：3，自引：0，他引：3
探讨黄瓜叶片氮代谢关键酶类活性与果实氮化物含量的相互关系,为黄瓜氮素的高效利用和品质优化提供依据。采用二因素裂区设计,研究了氮肥用量对黄瓜叶片氮代谢酶活性和结果盛期果实氮化物含量的影响。结果表明:在225~375 kg/hm~2施氮情况下,随施氮量增加功能叶片硝酸还原酶(NR)、谷氨酰胺合成酶(GS)、谷氨酸草酰乙酸转氨酶(GOT)和谷氨酸丙酮酸转氨酶(GPT)的活性逐渐升高,处理间差异显著,施氮量超过375 kg/hm~2其酶活性增加不明显且有下降的趋势。不同品种间酶活性大小依次为津优35号、园丰元6号、永昌9618,博特202,其中津优35号与园丰元6号酶活性差异不显著,与永昌9618和博特202差异显著。果实中氮化物的含量随施氮量的增加逐渐提高;施氮量为375 kg/hm~2时游离氨基酸、可溶性蛋白及粗蛋白含量达到最大值;增加到450 kg/hm~2时3种氮化物含量均下降,而硝酸盐含量持续增加;在525 kg/hm~2时含量最高,津优35号硝酸盐和游离氨基酸含量最低,可溶性蛋白和粗蛋白含量最高,博特202与之相反。相关分析表明,果实中硝酸盐含量与NR和GS活性呈显著正相关关系,与GPT和GOT无显著相关关系;可溶性蛋白和粗蛋白含量与4种酶类存在显著或极显著正相关关系。不同品种间果实氮化物的差异是氮素吸收同化综合作用的结果。选择氮素吸收同化能力强的品种,既有利于提高氮素利用,又可改善产品质量。 相似文献
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以农杂66为材料,研究了营养液漂浮育苗和营养钵育苗移栽棉生理生化特性的差异.结果表明:与营养钵育苗(对照)相比,营养液漂浮育苗有效地提高了移栽棉根系的TTC还原强度,棉株在苗期、蕾期、初花期、盛花期、铃期和吐絮期叶片的叶绿素含量分别比对照增加1.2%、19.2%、20.0%、10.6%、22.6%和13.6%,可溶性蛋白质含量分别比对照增加5.6%、12.3%、5.5%、8.7%、8.2%和4.1%,可溶性糖含量分别比对照增加3.1%、8.3%、8.1%、4.0%、3.2%和3.8%,叶片光合效率得到改善.在棉花整个生长发育时期,营养液漂浮育苗移栽棉叶片的超氧化物歧化酶(SOD)和过氧化物酶(POD)活性均明显高于对照,膜脂过氧化产物丙二醛(MDA)含量均低于对照(除苗期外).营养液漂浮育苗移栽棉叶片的游离脯氨酸含量逐渐增加,游离氨基酸含量高于对照,能有效改善棉株的生理代谢. 相似文献
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Xiang ZHANG Ming-yuan ZHOU Ya-bing LI Zhen-yu LIU Yuan CHEN De-hua CHEN 《农业科学学报》2021,20(5):1229-1238
Cotton bolls exhibit the lowest insecticidal efficacy among all organs of Bt cotton, which would ultimately affect the yield formation. The objective of this study was to investigate the effects of different urea concentrations on the seed Bt protein contents, seed cotton yield and the corresponding protein metabolism mechanism. The experiments were conducted during 2017–2018 cotton growing seasons. Two cultivars, Sikang 3(hybrid, SK3) and Sikang 1(conventional, SK1), were treated with six urea concentrations and their seed Bt protein contents were compared during boll formation period. The urea spray concentration had a significant effect on the seed Bt toxin content and seed cotton yield. Spraying of either 5 or 6% urea led to higher insecticidal protein contents and higher seed cotton yield for both cultivars. Moreover, the highest amino acid and soluble protein contents, as well as GPT and GOT activities, and lower protease and peptidase activities were observed at the 5 to 6% urea levels. Significant positive correlations between the seed Bt toxin and amino acid contents, and between the seed Bt toxin content and GPT activities were detected. The lower boll worm number and hazard boll rate were also observed with the 5 to 6% urea treatments, which may be the reason why nitrogen spraying increased the seed cotton yield. Therefore, our results suggested that the seed Bt toxin content and insect resistance were impacted markedly by external nitrogen application, and 5 to 6% urea had the greatest effect on insect resistance. 相似文献
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Enhancing boll protein synthesis and carbohydrate conversion by the application of exogenous amino acids at the peak flowering stage increased the boll Bt toxin concentration and lint yield in cotton 
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下载免费PDF全文 《农业科学学报》2023,22(6):1684-1694
In Bacillus thuringenesis (Bt) transgenic cotton, the cotton boll has the lowest insecticidal protein content when compared to the other organs. The present study investigated the effects of amino acid spray application at the peak flowering stage on the cotton boll Bt toxin concentration and yield formation. Boll protein synthesis and carbohydrate conversion were also studied to reveal the fundamental mechanism. Three treatments (i.e., CK, the untreated control; LA1, five amino acids; LA2, 21 amino acids) were applied to two Bt cultivars of G. hirsutum (i.e., the hybrid Sikang 3 and the conventional Sikang 1) in the cotton-growing seasons during 2017 and 2018. Amino acid spray application at the peak flowering stage resulted in an increase of 5.2–16.4% in the boll Bt protein concentration and an increase of 5.5–11.3% in the seed cotton yield, but there was no difference between the two amino acid treatments. In addition, amino acid applications led to increases in the amino acid content, soluble protein content, glutamate pyruvate transaminase (GPT) activity, glutamate oxaloacetate transaminase (GOT) activity, glucose content, fructose content and soluble acid invertase (SAI) activity. This study also found that Bt protein content, enhanced boll number and the weight of opened bolls were closely related to carbon and nitrogen metabolism. The Bt protein content had significant linear positive correlations with amino acid and soluble protein contents. Enhanced boll number had significant linear positive correlations with the GPT and GOT activities from 15–25 days after flowering (DAF). The weight of opened bolls from 55–65 DAF had a significant linear positive correlation with the SAI activity. These results indicate that the enhancement of boll protein synthesis and carbohydrate conversion by amino acid application resulted in a simultaneous increase in the boll Bt protein concentration and cotton lint yield. 相似文献
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转Bt基因抗虫棉33B的氮素代谢特征 总被引：5，自引：0，他引：5
以转B t基因抗虫棉新棉33B及其常规棉对照品种泗棉3号为材料,探讨转B t基因抗虫棉氮素代谢特征。结果显示:转基因抗虫棉33B主茎功能叶片中的全N含量、可溶性蛋白含量、硝酸还原酶(NR)活性以及主要氨基酸总量和游离氨含量均高于常规棉,其氮代谢十分旺盛;转基因抗虫棉33B的棉铃对位叶、铃壳中的全N含量亦高于常规棉,表明转基因抗虫棉33B具有较强的源、库生理优势。但在棉铃发育中后期,转基因抗虫棉33B营养物质向棉铃运输速度比常规棉低,从而导致单铃重减轻,衣分下降。 相似文献
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WANG Jun CHEN Yuan YAO Meng-hao LI Yuan WEN Yu-jin CHEN Yuan ZHANG Xiang CHEN De-hua 《农业科学学报》2015,14(10):1971-1979
Higher boll worm survival rates were detected after high temperature presented during square period in Bt cotton. The objective of this study was to investigate the effects of high temperature level on the Bt efficacy of two different types of Bt cotton cultivars at squaring stage. During the 2011 to 2013 cotton growth seasons, high temperature treatments ranged from 34 to 44°C in climate chambers, and field experiments under high temperature weather with various temperature levels were conducted to investigate the effects of the high temperature level on square Bt protein concentration and nitrogen metabolism. The climate chamber experiments showed that the square insecticidal protein contents reduced after 24 h elevated temperature treatments for both cultivars, whereas significant declines of the square insecticidal protein contents were detected at temperature 38°C, and only slightly numerical reductions were observed when temperature below 38°C. Similar high temperature responses were also observed at the two field experimental sites in 2013. Correspondingly, high temperature below 38°C seems have little effect on the square amino acid concentrations, soluble protein contents, glutamic-pyruvic transaminase(GPT) and glutamic-oxalacetic transaminase(GOT) activities as well as protease and peptidase activities; however, when the temperature was above 38°C, reduced soluble protein contents, enhanced amino acid concentrations, decreased GPT and GOT activities, bolstered protease and peptidase activities in square were detected. In general, the higher the temperature is(38°C), the larger the changes for the above compound contents and key enzymes activities of the square protein cycle. The findings indicated that the unstable insect resistance of the square was related to high temperature level during square stage. 相似文献
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Plant density is the cultivation practice usually employed to manipulate boll distribution, boll setting and yield in cotton production. In order to determine the effect of plant density on the insecticidal protein content of Bacillus thuringiensis(Bt) cotton plants, a study was conducted in Yangzhou University of China in 2015 and 2016. Five plant densities(PD1–PD5, representing 15 000, 30 000, 45 000, 60 000, and 75 000 plants ha–1) were imposed on two Bt cotton cultivars, Sikang 1(the conventional cultivar, SK-1) and Sikang 3(the hybrid cultivar, SK-3). The boll number per plant, boll weight and boll volume all decreased as plant density increased. As plant density increased from 15 000 to 75 000 plants ha~(–1), seed Bt protein content increased, with increases of 66.5% in SK-1 and 53.4% in SK-3 at 40 days after flowering(DAF) in 2015, and 36.8% in SK-1 and 38.6% in SK-3 in 2016. Nitrogen(N) metabolism was investigated to uncover the potential mechanism. The analysis of N metabolism showed enhanced soluble protein content, glutamic-pyruvic transaminase(GPT) and glutamate oxaloacetate transaminase(GOT) activities, but reduced free amino acid content, and protease and peptidase activities with increasing plant density. At 20 DAF, the seed Bt toxin amount was positively correlated with soluble protein level, with correlation coefficients of 0.825** in SK-1 and 0.926** in SK-3 in 2015, and 0.955** in SK-1 and 0.965** in SK-3 in 2016. In contrast, the seed Bt protein level was negatively correlated with free amino acid content, with correlation coefficients of –0.983** in SK-1 and –0.974** in SK-3 in 2015, and –0.996** in SK-1 and –0.986** in SK-3 in 2016. To further confirm the relationship of Bt protein content and N metabolism, the Bt protein content was found to be positively correlated with the activities of GPT and GOT, but negatively correlated with the activities of protease and peptidase. In conclusion, our present study indicated that high plant density elevated the amount of seed Bt protein, and this increase was associated with decreased boll number per plant, boll weight and boll volume. In addition, altered N metabolism also contributed to the increased Bt protein content under high plant density. 相似文献
16.
转Bt基因棉花不同生育期对土壤微生物的影响 总被引：4，自引：1，他引：4
采用转基因棉花晋棉26号和具有相似遗传背景的常规棉花品种晋棉7号进行大田试验,评价转基因棉花对土壤微生物的影响。试验结果表明,除吐絮期外,转基因棉花土壤细菌和真菌数量显著增加;土壤放线菌数量在蕾期、花期和吐絮期比对照显著提高,而在苗期显著降低,收获期差异不显著;土壤氨化细菌数量在蕾期、吐絮期、收获期显著提高,而在苗期和花期降低;固氮菌数量在苗期、花期、吐絮期、收获期显著提高,而在蕾期显著降低;土壤纤维菌数量在苗期、花期显著增加,而在蕾期和吐絮期显著降低,在收获期差异不明显。 相似文献