Key Laboratory of Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, 4 Xueyuan Road, 571101, Haikou, China.
Planta. 2013 Apr;237(4):1025-36. doi: 10.1007/s00425-012-1820-x. Epub 2012 Dec 6.
A full-length abscisic acid (ABA) senescence and ripening inducible gene named LcAsr was obtained from litchi. Bioinformatic analysis showed that full-length LcAsr was 1,177 bp and contained an open reading frame (ORF) encoding 153 amino acids, 85- and 146-bp 5' and 3' UTRs, respectively. LcAsr was expressed in all organs, with preferential expression in the flower and low levels in pulp. The expression level of LcAsr in postharvest uncovered fruit reached a maximum at 24 h after harvest. When the litchi fruit was covered with plastic film, the LcAsr expression level remained constant. LcASR protein localized in the nucleus. LcAsr was transformed in Arabidopsis thaliana L. (ecotype Columbia) and four transgenic lines were obtained. One line, 35S::LcAsrD, was selected for drought tolerance analysis and showed higher tolerance to drought than the control. The activities of superoxide dismutase, catalase, ascorbate peroxidase, and glutathione reductase were much higher in the transgenic line than the control under drought conditions. The levels of several ABA/stress-regulated genes were investigated. The transcript level of responsive to ABA (RAB18) remained constant and responsive to dehydration (RD29A) displayed a slight decrease in the Columbia line (Col). However, the transcript levels of LcAsr, RAB18, and RD29A were greatly enhanced in the transgenic 35S::LcAsrD. The transcript levels of KAT1, KAT2, and SKOR were also markedly decreased in the transgenic line. These results suggest an important role of LcAsr as a protective molecule for water deficit and help to understand the molecular mechanism of postharvest litchi fruit dehydration.
从荔枝中获得了全长脱落酸(ABA)衰老和成熟诱导基因 LcAsr。生物信息学分析表明,全长 LcAsr 长 1177bp,包含一个开放阅读框(ORF),编码 153 个氨基酸,分别为 85bp 和 146bp 的 5'和 3'UTR。LcAsr 在所有器官中表达,在花中表达偏好,在果肉中表达水平较低。在采后荔枝果实中,LcAsr 的表达水平在收获后 24h 达到最高。当荔枝果实用塑料薄膜覆盖时,LcAsr 的表达水平保持不变。LcASR 蛋白定位于细胞核。LcAsr 被转化到拟南芥(Col 生态型)中,获得了四个转基因株系。一个株系 35S::LcAsrD 被选择进行耐旱性分析,表现出比对照更高的耐旱性。在干旱条件下,转基因株系的超氧化物歧化酶、过氧化氢酶、抗坏血酸过氧化物酶和谷胱甘肽还原酶活性均高于对照。还研究了几个 ABA/胁迫调节基因的水平。响应 ABA 的基因(RAB18)的转录水平保持不变,响应脱水的基因(RD29A)在哥伦比亚线(Col)中略有下降。然而,LcAsr、RAB18 和 RD29A 的转录水平在转基因 35S::LcAsrD 中大大增强。KAT1、KAT2 和 SKOR 的转录水平在转基因株系中也明显降低。这些结果表明 LcAsr 作为一种对水分亏缺有保护作用的分子,有助于理解采后荔枝果实脱水的分子机制。