中国水稻科学 ›› 2018, Vol. 32 ›› Issue (5): 427-436.DOI: 10.16819/j.1001-7216.2018.7037
张占田, 孙雅菲, 艾昊, 罗闻真, 冯冰, 孙文献, 徐国华, 孙淑斌*()
收稿日期:
2017-03-30
修回日期:
2017-08-18
出版日期:
2018-09-10
发布日期:
2018-09-10
通讯作者:
孙淑斌
基金资助:
Zhantian ZHANG, Yafei SUN, Hao AI, Wenzhen LUO, Bing FENG, Wenxian SUN, Guohua XU, Shubin SUN*()
Received:
2017-03-30
Revised:
2017-08-18
Online:
2018-09-10
Published:
2018-09-10
Contact:
Shubin SUN
摘要:
【目的】水稻OsSHR2(LOC_Os03g31880)基因为拟南芥AtSHR的同源基因,与OsSHR1、OsSCR1和OsSCR2 同属于水稻GRAS转录因子家族。已有研究报道,转录因子基因SHR和SCR共同调控植物根系、叶片的发育,并参与各项生命活动。本研究旨在阐明OsSHR2在水稻中的时空表达特征及其在营养生长中的调控作用。【方法】通过生物信息学分析、表达模式分析、萌发动力学分析和水培实验验证该基因的功能。【结果】生物信息学分析发现OsSHR2、OsSHR1、OsSCR1和OsSCR2与拟南芥和其他物种的SHR亚家族和SCR亚家族成员具有很高的序列一致性;表达模式和pOsSHR2::GUS材料染色分析发现,OsSHR2在整个生长发育过程中的根系、叶片、维管组织和生殖器官中表达强烈,并集中在根尖的中柱、侧根原基和叶片及茎维管组织的中心表达,在野生型的地上部和根系中,OsSHR2受缺磷影响下调表达;对获得的OsSHR2的CRISPR-Cas9突变体osshr2进行种子萌发实验和水培实验,发现与野生型相比,osshr2的萌发时间延后,萌发率降低,在正常供磷和缺磷处理下,osshr2的地上部和根系长度显著小于野生型。【结论】OsSHR2在地上部和根系的发育、维管组织形成以及营养与生殖生长中具有重要作用,这为今后OsSHR2在分子育种等领域的应用奠定理论基础。
中图分类号:
张占田, 孙雅菲, 艾昊, 罗闻真, 冯冰, 孙文献, 徐国华, 孙淑斌. 水稻转录因子基因OsSHR2的表达特征及其在营养生长中的调控作用[J]. 中国水稻科学, 2018, 32(5): 427-436.
Zhantian ZHANG, Yafei SUN, Hao AI, Wenzhen LUO, Bing FENG, Wenxian SUN, Guohua XU, Shubin SUN. Expression Patterns and Regulation of Transcription Factor Gene OsSHR2 in Vegetative Growth in Rice[J]. Chinese Journal OF Rice Science, 2018, 32(5): 427-436.
图1 不同物种SHR和SCR同源基因的生物信息学分析 A–拟南芥和水稻的SHR和SCR同源基因氨基酸和核苷酸的序列一致性分析;B–通过基因家族分析网站InterPro(http://www.ebi.ac.uk/ interpro/)进行拟南芥与水稻的SHR和SCR同源基因GRAS保守结构域和保守基序VHIID的位置预测;C–不同物种SHR和SCR同源基因的进化树分析。
Fig. 1. Bioinformatics analysis of SHR and SCR homologous genes of different species. A, Sequence homology analysis of amino acids and nucleic acid of SHR and SCR homologous genes in Arabidopsis thaliana and rice; B, Prediction of the positions of SHR and SCR homologous GRAS conserved domains and conserved motifs in Arabidopsis thaliana and rice by InterPro (http://www.ebi.ac.uk/interpro/); C, Phylogenetic tree analysis of SHR and SCR homologous genes of different species.
图3 pOsSHR2::GUS材料的GUS染色鉴定 A–种子根;B–种子根及侧根;C–新叶;D–新叶的放大图;E–叶原基;F–叶舌;G–茎节;H–茎及叶鞘;I–茎;J–茎基部;K–颖壳及小穗轴;L–颖花;M–子房和柱头;N–胚(萌发后3 d)。A~E,J:水稻苗期;F,G,I,K~N:水稻灌浆期;H:分蘖期。A~J中标尺为2 mm;K~N中标尺为0.5 mm。
Fig. 3. Identification of OsSHR2 promoter-driven tissue-specific GUS staining. A, Seed root; B, Seed root and lateral root; C, Young leaf; D, The enlarged view of the young leaf; E, Leaf primordium; F, Ligule; G, Node; H, Stem and leaf sheath; I, Stem; J, Basal stem; K, Husk and rachilla; L, Spikelet; M, Ovary and stigma; N, Embryo(three days after germination). A-E, J: Seedling stage; F, G, I, K-N: Grain filling stage; H, Tilling stage. A-J, Bar=2 mm; K-N, Bar=0.5 mm.
图4 WT和突变体osshr2的萌发及水培下的表型 A–萌发表型(萌发后3 d、5 d和8 d),标尺为5 cm;B–水培表型(萌发后14 d),标尺为5 cm;C–材料地上部和根系长度统计(萌发后3 d、5 d、8 d和14 d);DAG–萌发后天数。
Fig. 4. Phenotype and statistics of seed germination and hydroponics experiments of WT and osshr2. A, Germination phenotype(3, 5 and 8 days after germination), bar=5 cm; B, Hydroponics phenotype(14 days after germination), bar=5 cm; C, Shoot and root length (3, 5, 8 and 14 days after germination); DAG, Day after germination.
图5 OsSHR1和OsSHR2在水稻正常供磷与缺磷条件下的相对表达量 A–OsSHR1和OsSHR2在水稻地下部的相对表达量;B–OsSHR1和OsSHR2在水稻地上部的相对表达量。
Fig. 5. Relative expression level of OsSHR1 and OsSHR2 under Pi-sufficient and Pi-deficient conditions in rice. A, The relative expression of OsSHR1 and OsSHR2 in the roots; B, The relative expression of OsSHR1 and OsSHR2 in the shoots.
图6 WT和突变体osshr2在正常供磷和缺磷条件下的表型 A和B–缺磷3 d;C和D–缺磷1周;E和F–缺磷2周;G–缺磷2周的表型, 标尺为20 cm。
Fig. 6. Phenotype of the wild type(WT) and osshr2 under Pi-sufficient and Pi-deficient conditions. A and B, Pi-deficient lasting three days; C and D, Pi-deficient lasting one week; E and F, Pi-deficient lasting two weeks; G, The phenotype of material under Pi-sufficient and Pi-deficient lasting two weeks, bar=20 cm. c410, osshr2-c410; c207, osshr2-c207.
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