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园艺学报 ›› 2016, Vol. 43 ›› Issue (10): 1916-1928.doi: 10.16420/j.issn.0513-353x.2016-0410

• 果树 • 上一篇    下一篇

菠萝生长素极性运输载体基因AcPINs和AcAUXs的分离与表达分析

李运合1,2,3,孙光明1,张红娜1,刘胜辉1,吴青松1,3,*   

  1. (1中国热带农业科学院南亚热带作物研究所,广东湛江 524091;2农业部热带果树生物学重点实验室,广东湛江 524091;3海南省菠萝种质创新与利用工程技术研究中心,广东湛江 524091)
  • 出版日期:2016-10-25 发布日期:2016-10-25

Isolation and Expression Analysis of the Genes Encoding Polar Auxin Transporter AcPINs and AcAUXs in Pineapple

LI Yun-he1,2,3,SUN Guang-ming1,ZHANG Hong-na1,LIU Sheng-hui1,and WU Qing-song1,3,*   

  1. (1South Subtropical Crop Research Institute,Chinese Academy of Tropical Agricultural Sciences,Zhanjiang,Guangdong 524091,China;2Key Laboratory of Tropical Fruit Biology,Ministry of Agriculture,Zhanjiang,Guangdong 524091,China;3Hainan Provincial Engineering Research Center for Pineapple Germplasm Innovation and Utilization,Zhanjiang,Guangdong 524091,China)
  • Online:2016-10-25 Published:2016-10-25

摘要:

乙烯已经被广泛应用于菠萝人工催花,但其分子机制不是很清楚。以菠萝(Ananas comosus L.‘Perola’)茎尖为材料,采用RT-PCR结合RACE方法得到3个生长素极性运输输出载体基因AcPIN1AcPIN2AcPIN3)和2个生长素极性运输输入载体基因(AcAUX1AcAUX2)的cDNA及基因组DNA全长。AcPIN1AcPIN2AcPIN3AcAUX1AcAUX2的cDNA全长分别为2 690、2 388、2 057、2 156和1 580 bp,其开放读码框长度分别为1 854、1 917、1 530、1 479和1 392 bp,分别编码617、638、509、492和463个氨基酸;其基因组DNA全长分别为3 602、3 208、4 204、5 457和2 436 bp,从起始密码子到终止密码子的长度分别为3 244、2 780、3 947、5 264和2 321 bp。氨基酸序列多重比对及系统发育树结果显示AcPINs和AcAUXs分别属于植物PINs和AUX/LAXs家族。荧光定量PCR结果表明,菠萝茎尖经200和1 200 mg · L-1乙烯利诱导处理后,AcPINs的表达上调较多,其中AcPIN2在处理后的早期(1 ~ 2 d)和后期(28 ~ 37 d)显著上调,另外两个PIN家族基因AcPIN1AcPIN3在处理后的大部分时间都明显提高。AcAUX1的上调表达量相对较少,且相对表达量显著提高也主要集中在处理初期(1 d)和后期(28 ~ 37 d),而AcAUX2的上调表达则只在处理后的前期(1、2、9 d)。研究结果表明,乙烯利诱导菠萝成花过程中,存在着生长素极性运输,且生长素极性输出在此过程中的作用可能更重要

关键词: 菠萝, 开花, 生长素输出载体, 生长素输入载体, 生长素极性运输

Abstract:

Exogenous ethylene has been widely used to induce pineapple flowering,but the molecular mechanism behind ethephon induction is still unclear. Three genes encoding auxin efflux carriers (designated as AcPIN1AcPIN2 and AcPIN3),and two genes encoding auxin influx carriers(designated  as AcAUX1 and AcAUX2)were isolated from the pineapple(Ananas comosus L.‘Perola’)shoot apex using RT-PCR and RACE. The full-length cDNA of AcPIN1AcPIN2AcPIN3AcAUX1 and AcAUX2 were 2 690,2 388,2 057,2 156 and 1 580 bp,with the open reading frames of 1 854,1 917,1 530,1 479 and 1 392 bp,which encode a putative protein of 617,638,509,492 and 463 amino acids,respectively. The genomic DNA sequences of AcPIN1AcPIN2AcPIN3AcAUX1 and AcAUX2 were 3 602,3 208,4 204,5 457 and 2 436 bp,with the lengths of 3 244,2 780,3 947,5 264 and 2 321 bp,respectively,from the start codon to the terminator codon. The results of a phylogenetic tree analysis indicated that AcPINs and the AcAUXs belong to plant PINs and AUXs/LAXs groups. Quantitative real-time PCR indicated that the relative expressions of AcPINs were up-regulated much more than AcAUXs after the treatment of 200 and 1 200 mg · L-1 ethephon. The expression of AcPIN2 was significantly increased at the former period(1–2 d)and the later period(28–37 d)after the treatment. In contrast,the expressions of AcPIN1 and AcPIN3 were significantly up-regulated at most time points after the treatment. On the other hand,the up-regulated expression of AcAUX1 was mainly on the former period(1 d)and later period(28–37 d),while the significantly up-regulated expression of AcAUX2 was only found at the former period(1,2 and 9 d after the treatment). The results showed that there were polar auxin transport during the flowering of pineapple,and auxin efflux might play a more important role during this process.

Key words: Ananas comosus, flowering, auxin efflux carrier, auxin influx carrier, polar auxin transport

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