Tomato is an important horticultural and economic crop cultivated worldwide.As Phytophthora infestans becomes a huge threat to tomato production,it is necessary to study the resistance mechanisms of tomato against P.i...Tomato is an important horticultural and economic crop cultivated worldwide.As Phytophthora infestans becomes a huge threat to tomato production,it is necessary to study the resistance mechanisms of tomato against P.infestans.Our previous research has found that miR482 might be involved in tomato–P.infestans interaction.In this study,miR482b precursor was cloned from Solanum pimpinellifolium“L3708”and miR482b was shown to decrease in abundance in tomato following P.infestans infection.Compared to wild-type tomato plants,tomato plants that overexpressed miR482b displayed more serious disease symptoms after P.infestans infection,with more necrotic cells,longer lesion diameters,and increased P.infestans abundance.Meanwhile,silencing of miR482b was performed by short tandem target mimic(STTM),resulting in enhancement of tomato resistance to P.infestans.Using miRNA and degradome data sets,NBS–LRR disease-resistance genes targeted by miR482b were validated.Negative correlation between the expression of miR482b and its target genes was found in all miR482b-overexpressing and-silencing tomato plants.Our results provide insight into tomato miR482b involved in the response to P.infestans infection,and demonstrate that miR482b–NBS–LRR is an important component in the network of tomato–P.infestans interaction.展开更多
大豆疫霉根腐病是由大豆疫霉菌(Phytophthora sojae)引起的危害大豆生长的严重病害。课题组前期研究表明具有P-loop结构域的GmPR10(Gene Bank accession no.FJ960440)和具有P-loop、Bet v1结构域的Gly m 4l(Gene Bank accession no.HQ91...大豆疫霉根腐病是由大豆疫霉菌(Phytophthora sojae)引起的危害大豆生长的严重病害。课题组前期研究表明具有P-loop结构域的GmPR10(Gene Bank accession no.FJ960440)和具有P-loop、Bet v1结构域的Gly m 4l(Gene Bank accession no.HQ913577.1)抑制大豆疫霉菌生长,并且过表达GmPR10和Gly m 4l的转基因大豆植株可以提高对大豆疫霉根腐病的抗性。为研究GmPR10和Gly m 4l抑菌机理,本研究利用点突变技术,获得了GmPR10的P-loop结构域突变体(Gly48/Thr48和Gly^51/Arg^51)、Gly m 4l的P-loop结构域突变体(Gly^49/Ile^49和Lys^55/Pro^55)、GmPR10和Gly m 4l的P-loop结构域以及Gly m 4l的Bet v1结构域缺失突变体,并纯化回收相应突变体蛋白,进行体外抑制大豆疫霉菌试验。结果表明,突变或缺失P-loop,Bet v1结构域的GmPR10和Gly m 4l失去了抑制大豆疫霉菌(Race 1)生长的能力,说明P-loop、Bet v 1结构域对GmPR10和Gly m 4l行使抑菌功能至关重要。展开更多
基金This work was supported by Grants from the NFSC(Nos.31471880 and 61472061).
文摘Tomato is an important horticultural and economic crop cultivated worldwide.As Phytophthora infestans becomes a huge threat to tomato production,it is necessary to study the resistance mechanisms of tomato against P.infestans.Our previous research has found that miR482 might be involved in tomato–P.infestans interaction.In this study,miR482b precursor was cloned from Solanum pimpinellifolium“L3708”and miR482b was shown to decrease in abundance in tomato following P.infestans infection.Compared to wild-type tomato plants,tomato plants that overexpressed miR482b displayed more serious disease symptoms after P.infestans infection,with more necrotic cells,longer lesion diameters,and increased P.infestans abundance.Meanwhile,silencing of miR482b was performed by short tandem target mimic(STTM),resulting in enhancement of tomato resistance to P.infestans.Using miRNA and degradome data sets,NBS–LRR disease-resistance genes targeted by miR482b were validated.Negative correlation between the expression of miR482b and its target genes was found in all miR482b-overexpressing and-silencing tomato plants.Our results provide insight into tomato miR482b involved in the response to P.infestans infection,and demonstrate that miR482b–NBS–LRR is an important component in the network of tomato–P.infestans interaction.
文摘大豆疫霉根腐病是由大豆疫霉菌(Phytophthora sojae)引起的危害大豆生长的严重病害。课题组前期研究表明具有P-loop结构域的GmPR10(Gene Bank accession no.FJ960440)和具有P-loop、Bet v1结构域的Gly m 4l(Gene Bank accession no.HQ913577.1)抑制大豆疫霉菌生长,并且过表达GmPR10和Gly m 4l的转基因大豆植株可以提高对大豆疫霉根腐病的抗性。为研究GmPR10和Gly m 4l抑菌机理,本研究利用点突变技术,获得了GmPR10的P-loop结构域突变体(Gly48/Thr48和Gly^51/Arg^51)、Gly m 4l的P-loop结构域突变体(Gly^49/Ile^49和Lys^55/Pro^55)、GmPR10和Gly m 4l的P-loop结构域以及Gly m 4l的Bet v1结构域缺失突变体,并纯化回收相应突变体蛋白,进行体外抑制大豆疫霉菌试验。结果表明,突变或缺失P-loop,Bet v1结构域的GmPR10和Gly m 4l失去了抑制大豆疫霉菌(Race 1)生长的能力,说明P-loop、Bet v 1结构域对GmPR10和Gly m 4l行使抑菌功能至关重要。
