Expression of gene specific to ethylene-induced ripeness to flower in Dutch iris bulbs

荷兰鸢尾球茎乙烯诱导开花成熟特异基因的表达

• 批准号: 10660033
• 负责人: DOI Motoaki
• 金额: $ 1.79万
• 依托单位: Osaka Prefecture University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10660033/
• 关键词: Dutch iris; juvenile phase; ripeness to flower; ethylene; gene expression; RT-PCR; 3'RACE; RT-PCT; アミノ酸配列

Using Poly-T primers, cDNAs were transcripted from RNAs extracted from the shoot tip of Dutch iris 'Blue Magic' bulbs, and they were amplified through PCR. Gene expression was analyzed by deferential display method, electrophoresing these PCR products on agarose gel. A PCR product (namely CG-01) was detected specifically in shoot tip of small-size bulbs (9g) without ethylene application, which were considered to be in juvenile phase and insensitive to inductive chilling. Investigation was focusted on this gene fragment.Before exposing bulbs to high temperature in 1999, expression of CG-01 was detected strongly in small-size bulbs and slightly in large-size bulbs (17g). However, the expression of CG-01 disappeared after exposing them to 100ppm ethylene for 24 hr. The flowering percentage of small-size and large-size bulbs after chilling treatment were 0% and 33%, respectively, if ethylene was not applied, indicating that all or most of bulbs were at juvenile phase. DNA sequence of CG-01 toward the 3' terminal was analyzed by a 3'RACE method suing bulbs after exposing them to high temperature. A PCR product which was amplified by the adaptor primer and a synthetic primer adupted to CG-01 sequence was obtained. However, sequence of this PCR product and that of CG-01 already known was not homologous. These results indicated that expression of CG-01 disappeared after summer even in small size bulbs at juvenile phase.

以荷兰鸢尾‘蓝魔术’鳞茎茎尖为材料，用Poly-T引物扩增出cDNA，并进行PCR扩增。差异显示法分析基因表达，将这些PCR产物在琼脂糖凝胶上电泳。在未施乙烯的小鳞茎（9 g）茎尖中检测到一个特异的PCR产物CG-01，该产物被认为处于幼球期，对诱导冷处理不敏感。在1999年高温处理前，CG-01基因在小鳞茎中有较强的表达，而在大鳞茎（17 g）中有微弱的表达。100 ppm乙烯处理24 h后，CG-01基因的表达消失，未处理的小鳞茎和大鳞茎的开花率分别为0%和33%，表明大部分或全部鳞茎处于幼期。将CG-01的3'末端的DNA序列通过3' RACE方法使用暴露于高温后的鳞茎进行分析。用该衔接引物和与CG-01序列相匹配的合成引物进行PCR扩增，获得了一条PCR产物。然而，该PCR产物的序列与已知的CG-01的序列不同源。这些结果表明，CG-01的表达消失后，即使在小尺寸的灯泡在幼年期。