拟南芥转录共激活子ANGUSTIFOLIA3(AN3)调控花的雄蕊的形成
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摘要
雄蕊是种子植物产生花粉的重要生殖器官,其是否正常发育关乎到植物的繁殖状况,并且会对农作物的产量造成影响。通过RT-PCR技术鉴定拟南芥转录共激活子ANGUSTIFOLIA3(AN3)的两个敲除突变体an3-1和an3-4;通过形态学检测发现,突变体an3-1和突变体an3-4的雄蕊较野生型雄蕊短,而雌蕊却无明显变化;通过构建AN3启动子GUS表达载体,对Pro-AN3-GUS植株的花组织进行染色,并观察,结果表明,AN3基因在拟南芥的种子胚、成熟的花粉、柱头、花瓣中均有表达。这个结果证明AN3能在拟南芥生殖生长期间在花器官等重要组织中表达,这个结果与an3-1和an3-4的雄蕊变短的结论一致。由此,我们得出结论:拟南芥转录共激活子AN3正向调控花的雄蕊的形成。
Stamens are important reproductive organs which produce pollen in seed plants,and their normal development is related to the reproductive status of plants and affects the yield of crops. Two knockout mutants an3-1 and an3-4 of Arabidopsis thaliana transcription coactivator ANGUSTIFOLIA3( AN3) were identified by RT-PCR. By morphological examination,the stamens of the an3-1 and an3-4 mutants were shorter than those of wild-type,whereas, the pistils were not changed significantly. By constructing AN3 promoter with GUS expression vector,the flower tissues of obtained Pro-AN3-GUS plants were stained and observed. The AN3 promoter in Arabidopsis seed embryos,mature pollen,stigma,petals was expressed. AN3 could be expressed in important tissues such as floral organs during Arabidopsis reproductive growth. These above results are consistent with the short stamens of an3-1 and an3-4 mutants. Thus,AN3 positively regulates the stamen development of A. thaliana.
Stamens are important reproductive organs which produce pollen in seed plants,and their normal development is related to the reproductive status of plants and affects the yield of crops. Two knockout mutants an3-1 and an3-4 of Arabidopsis thaliana transcription coactivator ANGUSTIFOLIA3( AN3) were identified by RT-PCR. By morphological examination,the stamens of the an3-1 and an3-4 mutants were shorter than those of wild-type,whereas, the pistils were not changed significantly. By constructing AN3 promoter with GUS expression vector,the flower tissues of obtained Pro-AN3-GUS plants were stained and observed. The AN3 promoter in Arabidopsis seed embryos,mature pollen,stigma,petals was expressed. AN3 could be expressed in important tissues such as floral organs during Arabidopsis reproductive growth. These above results are consistent with the short stamens of an3-1 and an3-4 mutants. Thus,AN3 positively regulates the stamen development of A. thaliana.
引文
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2.徐义流,张绍玲.花粉-雌蕊相互作用的分子基础[J].西北植物学报,2003,23(10):1800-1809.Xu Y L,Zhang S L.Molecular basis of pollen-pistil interaction[J].Acta Botanica Boreali-Occidentalia Sinica,2003,23(10):1800-1809.
3.姜茜.糖与p H影响蚕豆花粉萌发率的实验[J].生物学通报,2001,36(4):43.JIANG Xi.Effects of sugar and p H on germination rate of Vicia faba L.pollen[J].Bulletin of Biology,2001,36(4):43.
4.Horiguchi G,Kim G T,Tsukaya H.The transcription factor At GRF5 and the transcription coactivator AN3 regulate cell proliferation in leaf primordia of Arabidopsis thaliana[J].The Plant Journal,2005,43(1):68-78.
5.Horiguchi G,Nakayama H,Ishikawa N,et al.ANGUSTIFO-LIA3 plays roles in adaxial/abaxial patterning and growth in leaf morphogenesis[J].Plant Cell Physiology,2011,52(1):112-124.
6.Kanei M,Horiguchi G,Tsukaya H.Stable establishment of cotyledon identity during embryogenesis in Arabidopsis by ANGUSTIFOLIA3 and HANABA TARANU[J].Development,2012,139(13):2436-2446.
7.Meng L S,Wang Y B,Loake G J,et al.Seed embryo development is regulated via an AN3-MINI3 gene cascade[J].Frontiers in Plant Science,2016,7:1645.
8.Kim J H,Kende H.A transcriptional coactivator,At GIF1,is involved in regulating leaf growth and morphology in Arabidopsis[J].Proceedings of the National Academy of Sciences of the United States of America,2004,101(36):13374-13379.
9.Meng L S,Li Y Q,Liu M Q,et al.The Arabidopsis ANGUS-TIFOLIA3-YODA gene cascade induces anthocyanin accumulation by regulating sucrose levels[J].Frontiers in Plant Science,2016,7:1728.
10.Meng L S.Transcription coactivator Arabidopsis ANGUSTI-FOLIA3 modulates anthocyanin accumulation and light-induced root elongation through transrepression of C onstitutive P hotomorphogenic1[J].Plant,Cell&Environment,2015,38(4):838-851.
11.Meng L S,Yao S Q.Transcription co-activator Arabidopsis ANGUSTIFOLIA3(AN3)regulates water-use efficiency and drought tolerance by modulating stomatal density and improving root architecture by the transrepression of YODA(YDA)[J].Plant Biotechnology Journal,2015,13(7):893-902.
12.高亢,马惠斌,吴立柱,等.拟南芥At CCa P2基因缺失突变体的鉴定及表型观察[J].河北农业大学学报,2012,35(5):67-71.Gao K,Ma H B,Wu L Z,et al.Identification and phenotype of T-DNA insert At CCa P2 gene mutant in Arabidopsis thaliana[J].Journal of Agricultural University of Hebei,2012,35(5):67-71.
13.Meng L S,Wang Y B,Yao S Q,et al.Arabidopsis AINTEG-UMENTA mediates salt tolerance by trans-repressing SCABP8[J].Journal of Cell Science,2015,128(15):2919-2927.
14.Meng L S,Wang Z B,Yao S Q,et al.The ARF2-ANT-COR15A gene cascade regulates ABA signaling-mediated resistance of large seeds to drought in Arabidopsis[J].Journal of cell science,2015,128(21):3922-3932.
15.何卓娜,王双双,马红,等.胞外多肽激素基因CHAL/CLL1/CLL2在拟南芥雄蕊发育过程中发挥重要作用[J].植物生理学报,2016,52(2):167-176.He Z N,Wang S S,Ma H,et al.Peptide hormones CHAL,CLL1 and CLL2 are important for stamen development in Arabidopsis[J].Plant Physiology Journal,2016,52(2):167-176.
16.周鹊,贾琦石,杨仲南,等.拟南芥雄性不育突变体ms1521的基因定位[J].上海师范大学学报:自然科学版,2008,37(3):296-300.Zhou Q,Jia Q S,Yang Z N,et al.Genetic mapping of an Arabidopsis thaliana male sterile gene ms1521[J].Journal of Shanghai Normal University:Natural Sciences,2008,37(3):296-300.
17.成志鹏.拟南芥心皮发育基因MS1522的功能分析及雄性不育突变体ems1227的基因定位[D].上海:上海师范大学,2012.Cheng Z P.Function analysis of MS1522 gene involved in Arabidopsis carpel development and genetic mapping of Arabidopsis male sterile mutant ems1227[D].Shanghai:Shanghai Normal University,2012.