关键词: 花生, 温度诱导载脂蛋白, 基因克隆, 表达分析

Abstract: The temperature-induced lipocalins（TILs）is correlated with the stability of plant under stress environment of hot，cold，lighting，and oxidizing. This study intended to clone the TIL gene from peanut and analyze its function in response to temperature stress. A gene encoding TIL，designated as AhTIL1，was screened from the peanut cDNA library. The relative expressions of the gene in the different tissues of peanuts，at the different stages of seeds，and under the temperature stress were detected by quantitative PCR. The results of sequence analysis showed that the length of the ORF of AhTIL1 was 558 bp，encoding 185 amino acids. The bioinformatics analysis indicated that the predicted molecular mass of encoded protein was 21.4 kD，ad pI was 6.78. Sequence alignment displayed that the amino acids of AhTIL1 were in high homology with TIL of other species. The predicted results of candidate promoter showed that the candidate gene promoter contained regulatory elements associated with the light reaction such as ACE，Box4，G-box，GAG-motif etc.，and other related cis regulatory elements such as ABA，salicylic acid，gibberellin，anaerobic etc. Microarray and RNA-seq results showed that AhTIL1 expressed in roots，stems，leaves，flowers，peg，and seeds，the highest in the flowers，second highest in the stems，and the least in the roots. After the peg penetrating into soil，the expression level of AhTIL1 decreased immediately，and then increased gradually with the enlarging and maturing of seeds. Real time quantitative PCR result proved that the expression level of AhTIL1 increased after low or high temperature treatment for 3，6，12 and 24 h，however，decreased after 48 h. These results indicated that AhTIL1 may play an important role in peanut response to temperature stress.

Key words: peanut, temperature-induced lipocalin, gene cloning, expression analysis