转3α-HSD基因水稻后代降解胆固醇的研究
摘要
以转入降解甾类物质关键酶基因3α-HSD的水稻为试验材料,进行降解胆固醇能力的检测;研究转基因水稻在添加胆固醇后农艺性状及品质性状的变化,以及转基因水稻差异蛋白质组学分析,得出主要结果如下:
1.本试验通过对转基因水稻后代及对照降解胆固醇能力的研究表明:在不同浓度下,转基因水稻对胆固醇降解的总趋势基本一致,都是呈先增加后降低的趋势,而且均在分蘖时期达到最高降解率。转基因水稻对添加3mg/L胆固醇的降解效果最好,对2mg/L胆固醇的降解效果次之,对1mg/L胆固醇的降解效果最弱。转基因水稻6个株系的降解率均显著高于对照,其中株系3、5的降解效果最好。
2.胆固醇影响下转基因水稻后代农艺性状的研究表明:胆固醇浓度对农艺性状影响比较大,而且农艺性状的变异也比较复杂。胆固醇浓度为3mg/L时,植株表现为株高较高,剑叶长势好,分蘖数和有效分蘖最多,但结实率偏低。当胆固醇浓度低于3mg/L时,植株株型较小,但结实率、千粒重较高。当胆固醇浓度高于3mg/L时,植株株型较小,但结实率、千粒重偏低。转基因水稻6个株系的株高、穗长、剑叶长、千粒重都低于对照。
3.胆固醇影响下转基因水稻后代品质性状的研究表明:转基因水稻主要品质性状由于胆固醇的影响都发生了不同程度的变化。特别是转基因水稻各株系籽粒的谷甾醇含量随外界胆固醇浓度的增加而增加,但是均显著低于对照,这与3α-HSD基因的功能特性有关,是本试验所希望得到的。同时,由于胆固醇浓度的增加,籽粒中可溶性蛋白的含量也发生了变化,但除个别株系外,转基因水稻与对照间无显著差异。
4.根据双向电泳分析结果,从双向电泳凝胶上找出9个差异点,进行MALDI-TOF-MS/MS分析,共有6个蛋白得到鉴定。按其功能分为4类:第一类,参与光合反应;第二类,防卫蛋白;第三类,参与抗性反应;第四类,参与能量代谢。
In this paper, the3α-HSD transgenic rice was used as material to detect the capacity of degrading cholesterol ,to investigate quality traits and agronomic traits under cholesterol and analyze the differential proteomics of transgenic rice.The results were as follows:
1. Study on the capacity of the 3α-HSD transgenic rices to degrade cholesterol demonstrated that the trend of degrading cholesterol increased then decreased under the different density of cholesterol, all to the highest degradation rate in tillering phase. Transgenic rices had the best degradation effect to 3mg/L cholesterol, inferior to 2 mg/L cholesterol, the worst degradation effect to 1mg/L cholesterol.The transgenic rices had the best degradation effect than the control.The strain 3 and 5 had the best degradation effect than the other strains.
2. Under the different density of cholesterol the agronomic traits analysis showed the difference between the transgenic rice and the control, and the difference was also existed in different transgenic strains. When the density of cholesterol was 3mg/L ,the transgenic plants had the higher plant height, well growing of the flag leaf,and also had the most tiller number and the effective tillers,but lower seed setting rate.When the density of cholesterol was lower than 3mg/L,the plant was smaller,but both seed setting rate and weight of 1000-grains were higher. When the density of cholesterol was above 3mg/L, the plant was small,both seed setting rate and weight of 1000-grains were lower.The transgenic strains were inferior to the control in the plant height,ear length, flag leaf and weight of 1000-grains.
3. The results of the quality characters under the different density of cholesterol showed that the quality characters of transgenic rices changed with the different density of cholesterol. The sitosterol content of six transgenic strains increased with the augment of density of cholesterol ,but lower than the control. With the augment of density of cholesterol, the soluble protein in transgenic rice seed also changed.Except for few strains, there was no significant difference between transgenic rice and the control.
4. 6 of 9 differentially expressed proteins were identified by MALDI-TOF-MS and database searching.These proteins were classified into 4 categories according to their function. The first group participated in the photosynthetic reaction. The second group was defensin. The third group participated in the resistant reaction. The fourth group participated in the metabolism of material and energy.
1.本试验通过对转基因水稻后代及对照降解胆固醇能力的研究表明:在不同浓度下,转基因水稻对胆固醇降解的总趋势基本一致,都是呈先增加后降低的趋势,而且均在分蘖时期达到最高降解率。转基因水稻对添加3mg/L胆固醇的降解效果最好,对2mg/L胆固醇的降解效果次之,对1mg/L胆固醇的降解效果最弱。转基因水稻6个株系的降解率均显著高于对照,其中株系3、5的降解效果最好。
2.胆固醇影响下转基因水稻后代农艺性状的研究表明:胆固醇浓度对农艺性状影响比较大,而且农艺性状的变异也比较复杂。胆固醇浓度为3mg/L时,植株表现为株高较高,剑叶长势好,分蘖数和有效分蘖最多,但结实率偏低。当胆固醇浓度低于3mg/L时,植株株型较小,但结实率、千粒重较高。当胆固醇浓度高于3mg/L时,植株株型较小,但结实率、千粒重偏低。转基因水稻6个株系的株高、穗长、剑叶长、千粒重都低于对照。
3.胆固醇影响下转基因水稻后代品质性状的研究表明:转基因水稻主要品质性状由于胆固醇的影响都发生了不同程度的变化。特别是转基因水稻各株系籽粒的谷甾醇含量随外界胆固醇浓度的增加而增加,但是均显著低于对照,这与3α-HSD基因的功能特性有关,是本试验所希望得到的。同时,由于胆固醇浓度的增加,籽粒中可溶性蛋白的含量也发生了变化,但除个别株系外,转基因水稻与对照间无显著差异。
4.根据双向电泳分析结果,从双向电泳凝胶上找出9个差异点,进行MALDI-TOF-MS/MS分析,共有6个蛋白得到鉴定。按其功能分为4类:第一类,参与光合反应;第二类,防卫蛋白;第三类,参与抗性反应;第四类,参与能量代谢。
In this paper, the3α-HSD transgenic rice was used as material to detect the capacity of degrading cholesterol ,to investigate quality traits and agronomic traits under cholesterol and analyze the differential proteomics of transgenic rice.The results were as follows:
1. Study on the capacity of the 3α-HSD transgenic rices to degrade cholesterol demonstrated that the trend of degrading cholesterol increased then decreased under the different density of cholesterol, all to the highest degradation rate in tillering phase. Transgenic rices had the best degradation effect to 3mg/L cholesterol, inferior to 2 mg/L cholesterol, the worst degradation effect to 1mg/L cholesterol.The transgenic rices had the best degradation effect than the control.The strain 3 and 5 had the best degradation effect than the other strains.
2. Under the different density of cholesterol the agronomic traits analysis showed the difference between the transgenic rice and the control, and the difference was also existed in different transgenic strains. When the density of cholesterol was 3mg/L ,the transgenic plants had the higher plant height, well growing of the flag leaf,and also had the most tiller number and the effective tillers,but lower seed setting rate.When the density of cholesterol was lower than 3mg/L,the plant was smaller,but both seed setting rate and weight of 1000-grains were higher. When the density of cholesterol was above 3mg/L, the plant was small,both seed setting rate and weight of 1000-grains were lower.The transgenic strains were inferior to the control in the plant height,ear length, flag leaf and weight of 1000-grains.
3. The results of the quality characters under the different density of cholesterol showed that the quality characters of transgenic rices changed with the different density of cholesterol. The sitosterol content of six transgenic strains increased with the augment of density of cholesterol ,but lower than the control. With the augment of density of cholesterol, the soluble protein in transgenic rice seed also changed.Except for few strains, there was no significant difference between transgenic rice and the control.
4. 6 of 9 differentially expressed proteins were identified by MALDI-TOF-MS and database searching.These proteins were classified into 4 categories according to their function. The first group participated in the photosynthetic reaction. The second group was defensin. The third group participated in the resistant reaction. The fourth group participated in the metabolism of material and energy.
引文
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