In Texas, the freezes of 1951 and 1962 together killed 125,000 acres of citrus trees and the freeze of 1983 killed 40,000 acres. The low temperature is one of the most important abiotic stresses to be understood and manipulated molecularly. Cold hardiness is found in the deciduous citrus relative, trifoliate orange, which can withstand temperatures as low as -26 °C when it is cold acclimated. Exposure of the cold hardy trifoliate orange plants to temperature from 28 °C to -5 °C enabled us to isolate and characterize one novel citrus low temperature gene (clt) with two transcripts, called clt-a and clt-b from leaves and twigs. Clt-a was produced when plants were subjected to low temperatures (starting at 10 °C), while cltb was constitutively expressed. Both clt-a and clt-b have the same open reading frame of 165 nucleotides and encodes a small protein of 54 amino acid. However, clt-a has an additional 98 bp nucleotides at the 3'-untranslated region (UTR), which is absent in clt-b. Expression analysis using relative quantitative RT-PCR demonstrated that clt-a is expressed exclusively at low temperatures, while clt-b is expressed constitutively (expression verified from 2 °C to -5 °C). In the process of deacclimation from -1 °C to 28 °C, the clt-a transcript degraded dramatically after 2 °C and was completely absent at 28 °C, while the clt-b transcript remain stable. When the acclimated plant was taken from -1 °C to room temperature, the clt-a gene degraded within 2 hours. Moreover, when acclimated plant was continuously exposed at -1 °C for 20 days, both transcripts clt-a and clt-b remained stable. Involvement of alternative splicing in transcript stability will be discussed.
Adriana Robbins, Ying Jia, and Eliezer Louzada*
Ying Jia, Dianren Xia, and E.S. Louzada
A cDNA coding for a putative terpene synthase (Grtps) was isolated from `Rio Red' grapefruit (Citrus paradisi Macf.) mature fruit by differential display RT-PCR and the corresponding full-length cDNA and genomic clone were subsequently obtained. The isolated cDNA clone was 1644 bp in length encoding a protein of 548 amino acids with a predicted molecular mass of 64 kDa and of pI 5.38. The genomic clone was 3203 bp in length with 6 introns and 7 exons. This Grtps appears to be a sesquiterpene synthase based on molecular weight, genomic organization, and similarity with the other terpene synthases. Both RT-PCR and Northern blot expression analysis indicated that Grtps is not expressed in immature fruits, roots, or leaves, but only in mature fruits. Southern blot analysis of genomic DNA demonstrated that Grtps is one of the members in the family of terpene synthases.
Wen-ji Xu, Feng-yang Yu, Qing-xiang Jia, Gang-jun Luo, and Xiao-ying Bi
Wei Hu, Ju-Hua Liu, Xiao-Ying Yang, Jian-Bin Zhang, Cai-Hong Jia, Mei-Ying Li, Bi-Yu Xu, and Zhi-Qiang Jin
The banana, a typical climacteric fruit, undergoes a postharvest ripening process followed by a burst in ethylene production that signals the beginning of the climacteric period. Postharvest ripening plays an important role in improving the quality of the fruit as well as limiting its shelf life. To investigate the role of glutamate decarboxylase (GAD) in climacteric ethylene biosynthesis and fruit ripening in postharvest banana, a GAD gene was isolated from banana, designated MuGAD. Coincidently with climacteric ethylene production, MuGAD expression as well as the expression of the genes encoding the Musa 1-aminocyclopropane-1-carboxylate synthase (MaACS1) and Musa 1-aminocyclopropane-1-carboxylate oxidase (MaACO1) greatly increased during natural ripening and in ethylene-treated banana. Moreover, ethylene biosynthesis, ripening progress, and MuGAD, MaACS1, and MaACO1 expression were enhanced by exogenous ethylene application and inhibited by 1-methylcyclopropene (1-MCP). Taken together, our results suggested that MuGAD is involved in the fruit ripening process in postharvest banana.