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1Innovations and stepwise evolution of CBFs/DREB1s and their regulatory networks in angiosperms显示文摘The C-repeat binding factors/dehydrationresponsive element binding protein 1 s(CBFs/DREB1 s)have been identified as major regulators of cold acclimation in many angiosperm plants.However,their origin and evolutionary process associated to cold responsiveness are still lacking.By integrating multi-omics data of genomes,transcriptomes,and CBFs/DREB1 s genome-wide binding profiles,we unveil the origin and evolution of CBFs/DREB1 s and their regulatory network.Gene collinearity and phylogeny analyses show that CBF/DREB1 is an innovation evolved from tandem duplication-derived DREBⅢgene.A subsequent event of e-whole genome duplication led to two CBF/DREB1 archetypes(CladesⅠandⅡ)in ancient angiosperms.In contrast to cold-insensitivity of Clade I and their parent DREBⅢgenes,CladeⅡevolved a further innovation in cold-sensitive response and was stepwise expanded in eudicots and monocots by independent duplications.In geological time,the duplication events were mainly enriched around the Cretaceous-Paleogene(K-Pg)boundary and/or in the Late Cenozoic Ice Age,when the global average temperature significantly decreased.Consequently,the duplicated CBF/DREB1 genes contributed to the rewiring of CBFs/DREB1 s-regulatory network for cold tolerance.Altogether,our results highlight an origin and convergent evolution of CBFs/DREB1 s and their regulatory network probably for angiosperms adaptation to global cooling.Yuqi Nie Liangyu Guo Fuqiang Cui Yirong Shen Xiaoxue Ye Deyin Deng Shuo Wang Jianhua Zhu Wenwu Wu 2022Journal of Integrative Plant Biology2022,64,11:1
2Natural variation in STAYGREEN contributes to low-temperature tolerance in cucumber显示文摘Low-temperature(LT)stress threatens cucumber production globally;however,the molecular mechanisms underlying LT tolerance in cucumber remain largely unknown.Here,using a genome-wide association study(GWAS),we found a naturally occurring single nucleotide polymorphism(SNP)in the STAYGREEN(CsSGR)coding region at the gLTT5.1 locus associated with LT tolerance.Knockout mutants of CsSGR generated by clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated nuclease 9 exhibit enhanced LT tolerance,in particularly,increased chlorophyll(Chl)content and reduced reactive oxygen species(ROS)accumulation in response to LT.Moreover,the C-repeat Binding Factor 1(CsCBF1)transcription factor can directly activate the expression of CsSGR.We demonstrate that the LT-sensitive haplotype CsSGRHapA,but not the LT-tolerant haplotype CsSGR^(HapG)could interact with NON-YELLOW COLORING 1(CsNYC1)to mediate Chl degradation.Geographic distribution of the CsSGR haplotypes indicated that the CsSGR^(HapG)was selected in cucumber accessions from high latitudes,potentially contributing to LT tolerance during cucumber cold-adaptation in these regions.CsSGR mutants also showed enhanced tolerance to salinity,water deficit,and Pseudoperonospora cubensis,thus CsSGR is an elite target gene for breeding cucumber varieties with broad-spectrum stress tolerance.Collectively,our findings provide new insights into LT tolerance and will ultimately facilitate cucumber molecular breeding.Shaoyun Dong Caixia Li Haojie Tian Weiping Wang Xueyong Yang Diane MBeckles Xiaoping Liu Jiantao Guan Xingfang Gu Jiaqiang Sun Han Miao Shengping Zhang 2023Journal of Integrative Plant Biology2023,65,12:0
3橡胶树蛋白激酶HbBIN2基因的克隆、蛋白表达纯化及酶活分析显示文摘橡胶树是天然橡胶的唯一材料来源。寒害是橡胶树面临的主要自然灾害之一,严重限制了橡胶树的生长发育和种植区域分布,克隆和鉴定橡胶树低温应答基因尤为重要。蛋白激酶BIN2(brassinosteroid insensitive 2)是一种丝氨酸/苏氨酸蛋白激酶,在调控植物应对低温胁迫中发挥重要作用,但橡胶树蛋白激酶HbBIN2还未被克隆与鉴定。本研究从橡胶树cDNA中成功克隆出HbBIN2,序列分析表明:HbBIN2的开放阅读框为1143 bp,编码380个氨基酸,蛋白的分子量为42.9 kDa,理论等电点为8.74,是亲水性蛋白且无跨膜结构域。将HbBIN2构建到原核表达载体pET28a上,转化大肠杆菌BL21(DE3)表达菌株,摸索合适的诱导条件后成功诱导表达出HbBIN2蛋白。比较HbBIN2在不同的诱导温度(16、28、37℃)、诱导时间(3、6、12 h)和IPTG浓度(0.1、0.3、0.5 mmol/L)下的表达量,结果表明,HbBIN2在37℃,0.3 mmol/L IPTG诱导12 h的表达量最大。对HbBIN2蛋白体外纯化条件进行探索,结果表明,100 mmol/L咪唑能将目的蛋白完全洗脱。纯化HbBIN2蛋白并进行激酶活性鉴定,结果表明,该蛋白具有激酶活性。该研究为后续HbBIN2蛋白功能分析和橡胶树应对低温胁迫的调控机制研究提供参考,为橡胶树耐寒品种分子育种提供重要的基因资源。郭靖 袁红梅 2022热带作物学报2022,43,10:0
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