摘要
【目的】水稻的抽穗期对水稻地域适应性及水稻产量至关重要,对水稻抽穗期基因进行鉴定及功能分析,可以为水稻育种提供优异的基因资源。【方法】通过BSA-seq方法对一个黄化早抽穗突变体hz1(huangzao1)进行基因定位克隆及连锁分析;利用RT-qPCR技术分析目的基因HZ1的表达谱,并用水稻原生质体瞬时转化查明HZ1的亚细胞定位;对突变体的抽穗期、叶绿素含量、过氧化氢含量等生理指标进行测定,详细分析其表型变化。【结果】田间表型观察发现hz1表现早抽穗,长日照(long-day,LD)及短日照(short-day,SD)条件下hz1的抽穗期相同,分别比野生型(wild type,WT)早抽穗43 d和26 d,表明hz1是一个光周期不敏感的突变体。同时hz1表现黄化表型,相比WT,叶绿素含量下降。遗传分析表明hz1由隐性单基因控制,F2混池高通量测序将目的基因定位于水稻第6染色体上17.8 Mb区间内,分析发现该区间内一个T-DNA插入位点LOC_Os06g40080与hz1目标性状完全连锁,LOC_Os06g40080为已知的SE5基因,编码血红素加氧酶1(heme oxygenase 1,HO1)。HZ1/SE5在叶片中高表达,在LD及SD条件下具有昼夜节律性表达。亚细胞定位发现HZ1/SE5蛋白定位于叶绿体中。表达调控分析表明HZ1/SE5主要通过调控水稻成花素Hd3a和RFT1的表达来调控水稻的抽穗期;并通过调控叶绿素合成途径相关基因的表达水平影响水稻叶绿素水平变化。【结论】黄化早抽穗突变体hz1由于血红素加氧酶编码基因SE5突变导致其对光周期不敏感,HZ1/SE5基因通过调控水稻成花素基因及叶绿素合成途径相关基因的表达而影响水稻的抽穗期及叶片的黄化。
【Objective】The heading date of rice(Oryza sativa)is an important agronomic trait and crucial for the regional adaptability and yields.Identification and functional analysis of heading date-associated genes may provide important gene resources for the breeding in rice.【Method】BSA-seq was performed to identify target gene of a yellowish and early heading mutant huangzao 1(hz1).RT-qPCR was used to analyze the expression profile of target genes.The subcellular localization of HZ1 was determined by transient transformation between rice protoplasts and tobacco cells.Meanwhile,the heading dates,chlorophyll content and hydrogen peroxide content were measured to analyze the phenotype of hz1 mutant.【Result】Field phenotypic observations found that hz1 showed early heading phenotype.The heading date of hz1 mutant was the same under long-day(LD)and short-day(SD)conditions,which were 43 d and 26 d earlier than those of wild type(WT)respectively,indicating that hz1 was a photoperiod-insensitive mutant.Moreover,hz1 demonstrated a yellowish phenotype,with a decrease in chlorophyll content compared with WT.Genetic analysis revealed that hz1 was controlled by a recessive gene.BSA-seq(Bulk segregation analysis with whole-genome sequencing)of F2 pools indicated that the causal gene was linked with molecular markers on the chromosome 6 with 17.8 Mb region.Further analysis demonstrated that LOC_Os06g40080 site with a T-DNA insertion was completely linkage with hz1 mutant.LOC_Os06g40080 site was an identified SE5 gene encoding heme oxygenase 1(HO1).Expression pattern analysis revealed that HZ1/SE5 was highly expressed in the leaves and dhad diurnal rhythm expression.Subcellular localization assay showed that HZ1/SE5 protein was localized in chloroplasts.Gene expression analysis showed that HZ1/SE5 regulated the expression of florigen genes Hd3a and RFT1 to modulate the heading date in rice.In addition,HZ1/SE5 can also regulate the expression of genes associated with chlorophyll synthesis pathway to modulate the change of chl
作者
庞梦真
徐汉琴
刘海燕
宋娟
王佳涵
孙丽娜
姬佩梅
尹泽芝
胡又川
赵晓萌
梁闪闪
张泗举
栾维江
PANG Meng-zhen;XU Han-qin;LIU Hai-yan;SONG Juan;WANG Jia-han;SUN Li-na;JI Pei-mei;YIN Ze-zhi;HU You-chuan;ZHAO Xiao-meng;LIANG Shan-shan;ZHANG Si-ju;LUAN Wei-jiang(College of Life Sciences,Tianjin Normal University,Tianjin 300387;Tianjin Key Laboratory of Animal and Plant Resistance,Tianjin 300387)
出处
《生物技术通报》
CAS
CSCD
北大核心
2024年第7期125-136,共12页
Biotechnology Bulletin
基金
国家自然科学基金项目(31770343)
天津市水稻产业技术体系创新团队项目(ITTRRS2021100004)。
