Abstract:Abstract Siraitia Grosvenorii is an dioecious plant with medicinal and editable fruits as harvest, native to north of Guangxi Province, China. To explore the molecular mechanism of ethylene signal pathway, based on the unigene information from transcriptome data and combined with the rapid amplification of cDNA ends (RACE) technique and high-efficiency thermal asymmetric interlaced polymerase chain reaction (hi-TAIL PCR) technique, the full length mRNA and DNA of the key enzyme of ethylene synthesis was obtained from S. grosvenorii. The relative expression of genes on the male plant, female and Ag+treated female plants at different stages of different tissue parts were detected by real-time fluorescence quantitative technique. The results showed that the total length of the mRNA was 1 954 bp, named as SgACS3 (GenBank accession number: KY705404), and the full length DNA of the gene was 2 530 bp and contains three introns. The expressions of SgACS3 gene of the female leaves and buds before pollination were particularly high. The relative expressions of buds and leaves in the female plants were significantly higher than those in Ag+treated plants, suggesting that SgACS3 gene was widely involved in the growth and development of S. grosvenorii, stamen development in the buds, and it was closely related to the formation of female flowers and Ag+ by impact negatively regulate expression. This study provides an important base for revealing the molecular mechanism of SgACS3 in regulating the sex express and organ morphogenesis of flower and its application in breeding.
[1] 吕淑芳, 江静, 等. 2014.拟南芥乙烯合成酶ACS基因家族研究进展[J]. 生物技术通报,11:7-13.(LV Shu-Fang, JIANG Jing. 2014. Review of Arabidopisis 1-Aminocyclopropane-1-Carboxylic Acid Sythases[J].Biotechnology Bulletin, 11:7-13.)[2] 陈瑶瑶, 严良文, 刘智成, 等. 2014.苦瓜性别分化研究进展[J]. 福建农业科技, 45(9):69-72.(CHEN Yao-Yao, YAN Liang-Wen, LIU Zhi-Cheng, et al. 2014. Research progress in sex differentiation of bitter gourd (Monordica charantia L.)[J].FUJIAN AGRICULTRAL SCIENCE AND TECHNOLOGY, 45(9):69-72.)[3] 李锋, 蒋水元, 李典鹏.2010.罗汉果栽培与化学研究[M].广西科学技术出版社, 南宁.pp. 1-244.(LI Feng, JIANG Yong-Yuan, LI Dian-Peng. 2010. STUDY ON CULTIVATION AND CHEMISTRY OF SIRAITIA GROSVENORII[M]. Guangxi science and technology press, Nanning.pp.1-244.)[4] 杨龚, 唐燕琼. 2004. AgNO3、GA3和温度对苦瓜纯雌雄株系性别分化的影响[J]. 热带作物学报, 25(3):80-84.(YANG Gong, TANG Yan-Qiong. 2004. Eeffect of Chemical Application and Temperature on Sex Expression in Bitter Gourd (Monordica charantia L.)[J]. CHINESE JOURNAL OF TROPICAL CROPS, 25(3):80-84.)[5] 曾娜霞,胡姗姗,周琼,等. 2017. 罗汉果乙烯合成酶SgACS1基因的克隆与表达分析[J].分子植物育种,15(3): 821-832.(ZENG Na-Xia, HU Shan-Shan, Zhou Qiong, et al.2017. Cloning and Expression Analysis of 1-aminocyclopropane-1-carboxylic Acid Gene (SgACS1) of Siraitia grosvenorii[J]. Molecular Plant Breeding, 15(3):821-832.)[6] Abeles F B, Morgan P W, Saltveit ME Jr. 1992.Ethylene in plant biology, 2nd edn[M]. Academic Press, New York.PP.120-181.[7] Boualem A, Troadec C, Camps C, et al. 2015.A cucurbit androecy gene reveals how unisexual flowers develop and dioecy emerges[J]. Science, 350(6261): 688-691.[8] Chi-Kuang Wen. 2015.Ethylene in Plants[M]. Springer Science+Business Media, New York.pp.1-290.[9] Doyle J J, Doyle J L.1987.A rapid DNA isolation procedure for small quantities of fresh leaf tissue[J]. Phytochem Bull, 19: 11–15[10] Grichko V P, Glick B R. 2011.Amelioration of flooding stress by ACC deaminase-containing plant growth- promoting bacteria[J]. Plant Physiology and Biochemistry, 39: 11-17.[11] Liu Y G, Chen Y. 2007.High-efficiency thermal asymmetric interlaced PCR for amplification of unknown flanking sequences[J]. BioTechniques, 43:649-656.[12] Salman-Minkov A, Levi A, Wolf S, et al. 2008.ACC synthase genes are polymorphic in watermelon (Citrullus spp) and differentially expressed in flowers and in response to auxin and gibberellins[J]. Plant and Cell Physiology, 49(5): 740-750.[13] Tsuchisaka A, Theologis A. 2004.Unique and overlapping expression patterns among the Arabidopsis 1-amino-cyclopropane-1-carboxylate synthase gene family members[J]. Plant Physiology, 136(2) : 2982-3000[14] Ralph SG, Hudgins JW, Jancsik S, et al. 2007.Aminocyclopropane carboxylic acid synthase is a regulated step in ethylene-dependent induced conifer defense. Full-length cDNA cloning of a multigene family, differential constitutive, and wound-and insect-induced expression, and cellular and subcellular localization in spruce and Douglas fir[J]. Plant Physiol,143 (1):410–24.[15] von Dahl CC, Winz RA, Halitschke R, et al.2007. Tuning the herbivore-induced ethylene burst: the role of transcript accumulation and ethylene perception in Nicotiana attenuate[J]. Plant Journal,51 (2):293–307.[16] Varanasi V, Shin S, Mattheis J, et al. 2011.Expression profiles of the Md-ACS3 gene suggest a function as an accelerator of apple(Malus×domestica)fruit ripening[J]. Postharvest Biol Technol, 62(2):141-148.