TY - JOUR
T1 - Functional analyses of lipocalin proteins in tomato
AU - Wahyudi, Anung
AU - Ariyani, Dinni
AU - Ma, Gang
AU - Inaba, Ryosuke
AU - Fukasawa, Chikako
AU - Nakano, Ryohei
AU - Motohashi, Reiko
N1 - Funding Information:
We gratefully acknowledge Prof. Tsuyoshi Nakagawa (Shimane University) for providing pUGW5, pUGW6, and pGWB8 vectors to analyze the localization of proteins and to make over-expressed plants. We thank Prof. Yoshikazu Kiriiwa (Shizuoka University), Prof. Mitsuo Omura (Shizuoka University) and Prof. Takashi Nakatsuka (Shizuoka University) for valuable discussion on the technical aspects of this study. This work was supported by the Ministry of Education, Culture, Sports, Science and Technology (Japan) [Grants-in-Aid for Scientific Research (No. 23580039 to R M)] and Ministry of Research, Technology and Higher Education of the Republic of Indonesia, Directorate General of Resources for Research, Technology, and Higher Education.
Publisher Copyright:
© 2018 The Japanese Society for Plant Cell and Molecular Biology.
PY - 2018/12/25
Y1 - 2018/12/25
N2 - In this study, two temperature-induced lipocalin genes SlTIL1 and SlTIL2, and a chloroplastic lipocalin gene SlCHL were isolated from ‘Micro-Tom’ tomato. The coding sequences of SlTIL1, SlTIL2 and SlCHL were 558, 558, and 1002 bp, respectively. By TargetP analysis, no characteristic transit peptides were predicted in the proteins of SlTIL1 and SlTIL2, while a chloroplastic transit peptide was predicted in the protein of SlCHL. The subcellular localization results indicated that SlTIL1 and SlTIL2 proteins were major localized in the plasma membrane, while SlCHL was localized in chloroplast. To understand the function of lipocalins, transgenic tomato over-expressed SlTIL1, SlTIL2 and SlCHL and their virus-induced gene silencing (VIGS) plants were generated. The phenotypes were significantly affected when the SlTIL1, SlTIL2 and SlCHL were over-expressed or silenced by VIGS, which suggested that the three lipocalins played important roles in regulating the growth and development of tomato. In addition, the level of ROS (O − 2 and H 2 O 2 ) was low in SlTIL1, SlTIL2 and SlCHL over-expressed plants, while it was high in their silenced plants. The changes in the expression of SODs were consistent with the accumulations of ROS, which indicated that lipocalins might have an important role in abiotic oxidative stress tolerance in tomato plants. Especially SlTIL1 and SlTIL2 are localized around their membranes and protect them from ROS. The results will contribute to elucidating the functions of lipocalin in plants, and provide new strategies to improve the tolerance to abiotic stress in tomato plants.
AB - In this study, two temperature-induced lipocalin genes SlTIL1 and SlTIL2, and a chloroplastic lipocalin gene SlCHL were isolated from ‘Micro-Tom’ tomato. The coding sequences of SlTIL1, SlTIL2 and SlCHL were 558, 558, and 1002 bp, respectively. By TargetP analysis, no characteristic transit peptides were predicted in the proteins of SlTIL1 and SlTIL2, while a chloroplastic transit peptide was predicted in the protein of SlCHL. The subcellular localization results indicated that SlTIL1 and SlTIL2 proteins were major localized in the plasma membrane, while SlCHL was localized in chloroplast. To understand the function of lipocalins, transgenic tomato over-expressed SlTIL1, SlTIL2 and SlCHL and their virus-induced gene silencing (VIGS) plants were generated. The phenotypes were significantly affected when the SlTIL1, SlTIL2 and SlCHL were over-expressed or silenced by VIGS, which suggested that the three lipocalins played important roles in regulating the growth and development of tomato. In addition, the level of ROS (O − 2 and H 2 O 2 ) was low in SlTIL1, SlTIL2 and SlCHL over-expressed plants, while it was high in their silenced plants. The changes in the expression of SODs were consistent with the accumulations of ROS, which indicated that lipocalins might have an important role in abiotic oxidative stress tolerance in tomato plants. Especially SlTIL1 and SlTIL2 are localized around their membranes and protect them from ROS. The results will contribute to elucidating the functions of lipocalin in plants, and provide new strategies to improve the tolerance to abiotic stress in tomato plants.
KW - Lipocalins
KW - SlCHL
KW - SlTIL
KW - Tomato
KW - Virus-induced gene silencing
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U2 - 10.5511/plantbiotechnology.18.0620a
DO - 10.5511/plantbiotechnology.18.0620a
M3 - Article
AN - SCOPUS:85060844714
SN - 1342-4580
VL - 35
SP - 303
EP - 312
JO - Plant Biotechnology
JF - Plant Biotechnology
IS - 4
ER -