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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(AcPIN1、AcPIN2和AcPIN3)和2个生长素极性运输输入载体基因(AcAUX1和AcAUX2)的c DNA及基因组DNA全长。AcPIN1、AcPIN2、AcPIN3、AcAUX1和AcAUX2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AcPIN1和AcPIN3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。
Ethylene has been widely used in pineapple artificial flower, but its molecular mechanism is not very clear. Three auxin polar transport export vector genes (AcPIN1, AcPIN2 and AcPIN3) and two auxin polar transport inputs were obtained by RT-PCR combined with RACE using the shoot tip of Ananas comosus L.’Perola ’ Full length cDNA and genomic DNA of vector genes (AcAUX1 and AcAUX2). The full-length cDNAs of AcPIN1, AcPIN2, AcPIN3, AcAUX1 and AcAUX2 were 2690, 2388, 2557, 2156 and 1 580 bp, respectively. The open reading frames of them were 1 854, 1 917, 1 530 and 1 479 And 1 392 bp, encoding 617, 638, 509, 492 and 463 amino acids, respectively. The full-length genomic DNA was 3 602,3 208,4 204,5 457 and 2 436 bp, respectively. From the start codon to the stop codon The sub-lengths are 3 244, 2 780, 3 947, 5 264 and 2 321 bp, respectively. Multiple alignment of amino acid sequences and phylogenetic tree results showed that AcPINs and AcAUXs belonged to plant PINs and AUX / LAXs families, respectively. Quantitative real-time PCR results showed that the expression of AcPINs was significantly up-regulated after 200 and 1 200 mg · L -1 ethephon in pineapple shoot tips, and AcPIN2 up-regulated in the early (1-2 d) and later (28-37 d), and the other two PIN family genes, AcPIN1 and AcPIN3, were significantly increased after most of the treatment. AcAUX1 upregulated relatively less, and the relative expression increased significantly mainly in the early (1 d) and late (28 ~ 37 d), while AcAUX2 up-regulated expression only in the pre-treatment (1,2 , 9 d). The results showed that during the process of ethephon inducing pineapple into flower, the auxin polarity transport exists, and the role of auxin polar output in this process may be more important.