A cDNA Library was constructed with the heat shocked tomato ( Lycopersicon esculentum Mill.) flowers and then was screened with the probes of mitochondrial and endoplasmic reticulum conservative regions that were clon...A cDNA Library was constructed with the heat shocked tomato ( Lycopersicon esculentum Mill.) flowers and then was screened with the probes of mitochondrial and endoplasmic reticulum conservative regions that were cloned by using RT-PCR. The complete cDNAs of mitochondrial and endoplasmic reticulum small heat shock protein ( shsp) were selected out from the cDNA library. Furthermore, the temperature responses of these shsp genes were determined. Northern hybridization showed that the heat response temperatures of both genes in tomato flower were lower than that in leaf and that mitochondria shsp in leaf was cold-inducible. In this paper, the molecular features of the cloned genes, the causes of the uncommon heat response temperatures of sHSP in newer and the cold inducible character of mitochondria shsp gene in leaf were discussed.展开更多
Soybean(Glycine max(L.)Merr.)is an important cultivated crop,which requires much water during its growth,and drought seriously affects soybean yields.Studies have shown that the expression of small heat shock proteins...Soybean(Glycine max(L.)Merr.)is an important cultivated crop,which requires much water during its growth,and drought seriously affects soybean yields.Studies have shown that the expression of small heat shock proteins can enhance drought resistance,cold resistance and salt resistance of plants.In this experiment,soybean GmHsps_p23-like gene was successfully cloned by RT-PCR,the protein encoded by the GmHsps_p23-like gene was subjected to bioinformatics analysis,and the pCAMBIA3301-GmHsps_p23-like overexpression vector and pCBSG015-GmHsps_p23-like gene editing vector were constructed.Agrobacterium-mediated method was used to transform soybeans to obtain positive plants.RT-PCR detection,rehydration experiment and drought resistance physiological and biochemical index detection were performed on the T2 generation positive transgenic soybean plants identified by PCR and Southern hybridization.The results showed that the overexpression vector plant GmHsps_p23-like gene expression increased.After rehydration,the transgenic overexpression plants returned to normal growth,and the damage to the plants was low.After drought stress,the SOD and POD activities and the PRO content of the transgenic overexpression plants increased,while the MDA content decreased.The reverse was true for soybean plants with genetically modified editing vectors.The drought resistance of the overexpressed soybeans under drought stress was higher than that of the control group,and had a stronger drought resistance.It showed that the expression of soybean GmHsps_p23-like gene can improve the drought resistance of soybean.The cloning and functional verification of soybean GmHsps_p23-like gene had not been reported yet.This is the first time that PCR technology has been used to amplify the soybean GmHsps_p23-like gene and construct an expression vector for this gene.This research has laid the foundation for transgenic technology to improve plant drought resistance and cultivate new drought-resistant transgenic soybean varieties.展开更多
植物热激蛋白是一类代表性高温响应蛋白。以玉米种质POB21为材料,克隆了一个CDS序列长度为477 bp的小分子热激蛋白基因。该基因编码的蛋白含158个氨基酸,具有HSP20蛋白典型的ACD结构域,预测的等电点为5.36,分子量为17.746 k D,被命名为Z...植物热激蛋白是一类代表性高温响应蛋白。以玉米种质POB21为材料,克隆了一个CDS序列长度为477 bp的小分子热激蛋白基因。该基因编码的蛋白含158个氨基酸,具有HSP20蛋白典型的ACD结构域,预测的等电点为5.36,分子量为17.746 k D,被命名为Zm HSP17.7。在水稻、拟南芥等植物基因组中都有其同源基因,进化和亚细胞定位分析表明,此基因属CI类小分子热激蛋白家族成员。Northern杂交分析表明,高温快速诱导Zm HSP17.7基因表达,15%PEG模拟干旱胁迫不诱导该基因表达,但在复合胁迫下干旱增强了高温的诱导效果。外源ABA也不影响该基因的表达。与野生型拟南芥相比,超表达Zm HSP17.7的转基因拟南芥在种子萌发和植株生长过程中表现出更强的高温和干旱耐受性,说明该基因可以在植物防御高温、干旱及复合胁迫中发挥一定作用。展开更多
文摘A cDNA Library was constructed with the heat shocked tomato ( Lycopersicon esculentum Mill.) flowers and then was screened with the probes of mitochondrial and endoplasmic reticulum conservative regions that were cloned by using RT-PCR. The complete cDNAs of mitochondrial and endoplasmic reticulum small heat shock protein ( shsp) were selected out from the cDNA library. Furthermore, the temperature responses of these shsp genes were determined. Northern hybridization showed that the heat response temperatures of both genes in tomato flower were lower than that in leaf and that mitochondria shsp in leaf was cold-inducible. In this paper, the molecular features of the cloned genes, the causes of the uncommon heat response temperatures of sHSP in newer and the cold inducible character of mitochondria shsp gene in leaf were discussed.
基金Jilin Province Education Department Science and Technology Research Project[JJKH20210350KJ]Jilin Province Science and Technology Guidance Program Project[20200402023NC]+1 种基金Jilin Provincial Natural Science Foundation Project[20200201027JC]Innovation and Entrepreneurship Training Program for College Students in Jilin Province[2021].
文摘Soybean(Glycine max(L.)Merr.)is an important cultivated crop,which requires much water during its growth,and drought seriously affects soybean yields.Studies have shown that the expression of small heat shock proteins can enhance drought resistance,cold resistance and salt resistance of plants.In this experiment,soybean GmHsps_p23-like gene was successfully cloned by RT-PCR,the protein encoded by the GmHsps_p23-like gene was subjected to bioinformatics analysis,and the pCAMBIA3301-GmHsps_p23-like overexpression vector and pCBSG015-GmHsps_p23-like gene editing vector were constructed.Agrobacterium-mediated method was used to transform soybeans to obtain positive plants.RT-PCR detection,rehydration experiment and drought resistance physiological and biochemical index detection were performed on the T2 generation positive transgenic soybean plants identified by PCR and Southern hybridization.The results showed that the overexpression vector plant GmHsps_p23-like gene expression increased.After rehydration,the transgenic overexpression plants returned to normal growth,and the damage to the plants was low.After drought stress,the SOD and POD activities and the PRO content of the transgenic overexpression plants increased,while the MDA content decreased.The reverse was true for soybean plants with genetically modified editing vectors.The drought resistance of the overexpressed soybeans under drought stress was higher than that of the control group,and had a stronger drought resistance.It showed that the expression of soybean GmHsps_p23-like gene can improve the drought resistance of soybean.The cloning and functional verification of soybean GmHsps_p23-like gene had not been reported yet.This is the first time that PCR technology has been used to amplify the soybean GmHsps_p23-like gene and construct an expression vector for this gene.This research has laid the foundation for transgenic technology to improve plant drought resistance and cultivate new drought-resistant transgenic soybean varieties.
