赤霉素GA4是水稻矮化特征的重要调节因子

doi:10.3864/j.issn.0578-1752.2019.05.002

Abstract

Abstract:

【Objective】A dwarf rice mutant was generated by culturing rice embryo tissues and characterized to elucidate the reasons for leading to an occurence of semi-dwarf rice with more tiller number. It is expected that this study will provide a theoretical basis for scientifically cultivating rice varieties of lodging-resistant and high-yielding in overcoming the dwarf obstacle factors.【Method】In this study, both the rice dwarf mutant and wild type were phenotypically profiled, and the structural characteristics of flower and cell appearance of leaves were investigated by stereomicroscope and light microscopy; The differential gene expression profiles were analyzed by both the transcriptomics and qRT-PCR, and the sensitivity of the mutant to exogenous gibberellin was detected by spraying exogenous GA3; The enrichment profiles of gibberellin in the mutant were detected by a high performance liquid chromatography and a mass spectrometry. 【Result】Data showed that the mutant demonstrated a decrease of 56.59% in the average plant height and an increase of 47.44% in the effective tiller number compared with the wild type, respectively (P <0.01). Observation showed that the mutant led to disappearance of the epidermis and revealed a smaller stamen accompanying a delayed development in the flower organs. Although the mutant has a higher effective tiller number, but significantly lowers the seed setting rate, which only accounts for 12.62% of that in the wild type. The length and the width of grains also are significantly reduced (P <0.01). Stem longitudinal sections reveal that the mutant decreased the cell length of 23% compared with the wild type (P <0.01). However, when the mutant was exposed to exogenous gibberellin, the plant height, effective tiller number, seed setting rate, seed size, epidermis and the stem cell length were obviously restored, indicating that the dwarf mutant possibly results from the shortage of GA’s synthesis in plant. Transcriptome sequencing showed that the mutant significantly up-regulated the OsGA13ox gene, and exhibited an identical result with the qRT-PCR analyses. Since the OsGA13ox controls the conversion of the GA12 to the GA53, both of which are converted to the GA4 and the GA1, respectively. Particularly, the GA4 exhibits a higher activity than the GA1, suggesting that rice dwarf might be triggered by the reduction of GA4 enrichment in plant. Measurement confirmed that the accumulation of the GA4 in the mutant was decreased by 94.9% compared with the wild type. Additionally, as a specific receptor for SL (Strigolactone), the D14 gene is involved in the SL signaling transduction in plant, and thus inhibits branching or tillering. The qRT-PCR showed that the mutant significantly down-regulated the D14 gene compared to the wild type, however, both the wild type and the mutant significantly up-regulated after spraying GA3. The data suggested that the up-regulation of the D14 gene might lower the effective tiller number, while the down-regulation of the D14 gene possibly increase the effective tiller number. Statistical analyses demonstrates that the mutant significantly increased the effective tiller number, but both the wild type and mutant decreased the effective tiller number after spraying GA3, indicating that the expression profiles of the D14 gene in rice might be modulated by GA, and thereby exert on the tiller number.【Conclusion】Semi-dwarfed rice mutant is likely caused by a decrease of GA4 enrichment because of abnormal expression of the OsGA13ox gene, and GA might indirectly regulate rice tiller by affecting the expression of the D14 gene.

Key words: rice, semi-dwarf mutant, OsGA13ox, GA4, D14, effective tiller

HUANG ShengCai,WANG Bing,XIE GuoQiang,LIU ZhongLai,ZHANG MeiJuan,ZHANG ShuQing,CHENG XianGuo. Enrichment Profile of GA4 is an Important Regulatory Factor Triggering Rice Dwarf[J].Scientia Agricultura Sinica, 2019, 52(5): 786-800.

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基因 Gene	引物序列 Primer sequence (5′-3′)
β-Actin	F: CTGACAGGATGAGCAAGGAG R: GGCAATCCACATCTGCTGGA
OsGA13ox	F: AGAAGTGGAGAAAAGACTACGG R: CAATGATCTTTCTCTGGTGTGC
D14	F: AGAAAGAGAGAGAAGAAGCGAG R: CGCGCTCCCCTTTTATATACTA

基因 Gene	功能 Function
Os01g0115600	蛋白激酶功能,结合ATP Kinase function, binding with ATP
Os01g0781700	ATP结合功能ATP binding
Os10g0463800	功能未知Unknown function
Os03g0332100	GA13ox,催化GA12转化为GA53,决定GA4和GA1的比例^[24] GA13ox, Converts GA12 into GA53, determines the ratio of GA4 and GA1^[24]
Os04g0223500	黄素腺嘌呤二核苷酸,单加氧酶活性 FAD, monooxygenase activity
Os04g0606000	酰基转移酶活性 Transferase activity, transferring acyl groups
Os06g0364500	多糖结合功能Polysaccharide binding
Os12g0231700	推测是转位子蛋白Transposon protein, putative

基因 Gene	功能 Function
Novel00727	Lnc RNA,可能参与调控植物的表型Lnc RNA, probably participate in regulating the phenotype of plants
Os01g0191200	磷酸激酶活性 Acid phosphatase activity
Os01g0198900	功能未知 Unknow function
Os01g0368900	谷胱甘肽-二硫化物氧化还原酶活性 Has a glutathione-disulfide oxidoreductase activity
Os01g0501800	参与光合系统Ⅱ的组装和稳定 Involved in photosystem II assembly and stabilization
Os01g0600900	光受体,捕捉光信号 The light-harvesting complex functions as a light receptor
Os01g0948200	转录因子 DNA-binding transcription factor activity
Os03g0159900	功能未知 Unknow function
Os03g0196250	功能未知 Unknow function
Os03g0203200	D14蛋白参与独脚金内酯（SL）信号通路;与赤霉素竞争DELLA结合位点^[27] Involved in strigolactone signaling pathway; Competition with GA for DELLA binding sites^[27]
Os03g0251350	转录因子,调控长日照开花^[28]Probable transcription factor involved in the regulation of flowering time under long day^[28]
Os03g0856500	与核糖体小亚基结合,负调控蛋白翻译 Ribosomal small subunit binding, negative regulation of translational elongation

基因 Gene	基序 Motif	家族 Family	功能 Function
Os03g0224200	CAGTATCTTT	ARR-B	参与细胞分裂素信号的转导^[39] Participation in the transduction of cytokinin signaling^[39]
Os06g0130600	CACTCACATTCTCTCTGCACA	BBR-BPC	调控大麦BKn3 的表达^[40] Regulate the expression of BKn3 gene in barley^[40]
Os10g0419200	TTTTTTTTTTTG	C2H2	促进植物营养生长到生殖生长的转变^[41] Promote the transformation of plant vegetative growth to reproductive growth^[41]
Os07g0236700	GTATCTTTTTTTTTTTGCGCC	Dof^[42]	可能参与碳水化合物代谢基因的光调节,植物防御机制 Possible involvement in light regulation of carbohydrate metabolism genes, plant defense mechanisms
Os03g0821200	AACAAAGCTAAAACATAAGGG	Dof^[42]	参与种子萌发,发芽后糊粉中的赤霉素反应等基因的调控 Involving in seed germination, regulation of gibberellin reactions in aleurone after germination
Os03g0324300	GTCTAGATTCATTAC	EIL	乙烯信号响应的正向调控因子^[43] Positive regulator of ethylene signal response^[43]
Os03g0324200	GTCTAGATTCATTAC	EIL	在植物受伤时,参与乙烯信号的转导^[44] Participation in the transduction of ethylene signals when plants are injured^[44]
Os09g0490200	TAATGAATCTA	EIL	参与乙烯信号转导 Participation in ethylene signal transduction
Os11g0681200	GAAACACGCGCGCGC	FAR1	参与抵抗疾病^[45] Enhance the resistance of disease^[45]
Os02g0680700	TAGATATTA	MYB_related^[46]	功能未知 Unknown function
Os08g0157600	TAGATATTA	MYB_related^[46]	功能未知 Unknown function

Enrichment Profile of GA4 is an Important Regulatory Factor Triggering Rice Dwarf

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