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小麦中的热:利用反向遗传学技术在小分子热激蛋白26(sHsp26)家族中发现新的等位基因。

Heat in Wheat: Exploit Reverse Genetic Techniques to Discover New Alleles Within the sHsp26 Family.

作者信息

Comastri Alessia, Janni Michela, Simmonds James, Uauy Cristobal, Pignone Domenico, Nguyen Henry T, Marmiroli Nelson

机构信息

Department of Chemistry, Life Sciences and Environmental Sustainability, University of Parma, Parma, Italy.

Department of DiSBA, CNR, Institute of Bioscience and Bioresources, Bari, Italy.

出版信息

Front Plant Sci. 2018 Sep 19;9:1337. doi: 10.3389/fpls.2018.01337. eCollection 2018.

Abstract

Wheat breeding nowadays must address producers and consumers' desire. According to the last FAO report, a dramatic decrease in wheat production is expected in the next decades mainly due to the upcoming climate change. The identification of the processes which are triggered by heat stress and how thermotolerance develops in wheat is an active research topic. Genomic approach may help wheat breeding since it allows direct study on the genotype and relationship with the phenotype. Here the isolation and characterization of four members of the chloroplast-localized small heat shock proteins (sHSP) encoded by the gene family is reported. Furthermore, two high throughput TILLING (Targeting Induced Local Lesions In Genomes) approaches and were used for the identification of new alleles within this family. Small heat shock proteins are known to prevent the irreversible aggregation of misfolded proteins and contribute to the acquisition of thermotolerance. Chloroplast-localized sHSPs protect the photosynthetic machinery during episodes of high temperature stress. The modulation of the newly discovered genes within the sHsp26 family has been analyzed and by the ExpVIP platform widening the abiotic stress analysis; and their involvement in the heat stress response has been demonstrated. In addition, in this study a total of 50 TILLING mutant lines have been identified. A set of KASP (Kompetitive Allele Specific PCR) markers was also developed to follow the specific mutations in the ongoing backcrosses, applicable to high throughput genotyping approaches and usable in marker assisted selection breeding programs.

摘要

如今的小麦育种必须满足生产者和消费者的需求。根据粮农组织的最新报告,预计在未来几十年里,小麦产量将大幅下降,主要原因是即将到来的气候变化。确定热应激引发的过程以及小麦耐热性如何形成是一个活跃的研究课题。基因组方法可能有助于小麦育种,因为它允许直接研究基因型及其与表型的关系。本文报道了由该基因家族编码的四个叶绿体定位的小热激蛋白(sHSP)成员的分离和特性分析。此外,还使用了两种高通量定向诱导基因组局部突变(TILLING)方法来鉴定该家族中的新等位基因。已知小热激蛋白可防止错误折叠蛋白质的不可逆聚集,并有助于获得耐热性。叶绿体定位的sHSP在高温胁迫期间保护光合机制。通过ExpVIP平台对sHsp26家族中新发现基因的调控进行了分析,拓宽了非生物胁迫分析范围;并证明了它们参与热应激反应。此外,在本研究中总共鉴定出50个TILLING突变系。还开发了一组竞争性等位基因特异性PCR(KASP)标记,以追踪正在进行的回交中的特定突变,适用于高通量基因分型方法,并可用于标记辅助选择育种计划。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9b6f/6156267/dadd0e74afcb/fpls-09-01337-g0001.jpg

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