Identification of reference genes via real-time quantitative PCR for investigation of the transcriptomic basis of the squalene biosynthesis in different tissues on olives under drought stress

Squalene plays a crucial role in plant growth, development, and stress tolerance. To select the best reference genes (RGs) for expression profile analysis of genes involved in squalene biosynthesis in olives under stress tolerance, the expression stability of 22 candidate RGs across four tissues (ro...

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Main Authors: Haiyang Chi, Shasha Bai, Chenxiao Chen, Yuying Bao, Xiaoxin Qu, Shitao Sun, Jiangpeng Pan, Xiushi Yang, Chunsheng Hou, Yanchun Deng
Format: Article
Language:English
Published: Elsevier 2024-12-01
Series:Plant Stress
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Online Access:http://www.sciencedirect.com/science/article/pii/S2667064X24002288
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author Haiyang Chi
Shasha Bai
Chenxiao Chen
Yuying Bao
Xiaoxin Qu
Shitao Sun
Jiangpeng Pan
Xiushi Yang
Chunsheng Hou
Yanchun Deng
author_facet Haiyang Chi
Shasha Bai
Chenxiao Chen
Yuying Bao
Xiaoxin Qu
Shitao Sun
Jiangpeng Pan
Xiushi Yang
Chunsheng Hou
Yanchun Deng
author_sort Haiyang Chi
collection DOAJ
description Squalene plays a crucial role in plant growth, development, and stress tolerance. To select the best reference genes (RGs) for expression profile analysis of genes involved in squalene biosynthesis in olives under stress tolerance, the expression stability of 22 candidate RGs across four tissues (root, stem, leaf, and fruit) and five representative cultivars under drought stress was assessed using six methods. Our study showed that, ubiquitin-conjugating enzyme1 (UBC1) and 60S ribosomal protein L18-2-like (60S) were the most stable RGs across five different cultivars and various tissues. Additionally, elongation factor 1-alpha-like (EF1-α2) and UBC1 were the most stable RG in olives under drought stress. UBC1 was the appropriate RG for further study, qPCR analysis showed that five certain genes involved in the squalene biosynthesis exhibited significant differential expression under varying conditions. Specially, the expression level of squalene synthase (SQS) in the leaves was highest, and HPLC analysis showed that squalene content was the highest in leaves. Likewise, the expression levels of SQS and farnesyl diphosphate synthase (FPPS) in the leaves of the cultivar Arbosana and Chelmsford Lal were significantly higher than those of the other cultivars, respectively, and HPLC analysis also showed that squalene content was highest in the two cultivars. Interestingly, the amount of squalene increased dramatically in olive under drought stress, as lupeol synthase (LUPS) was significantly up-regulated. To date, this study firstly provided the comprehensive analysis of RGs related to squalene biosynthesis in various cultivars and tissues of olives under drought stress.
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publisher Elsevier
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spelling doaj-art-c36af6ab1c594646a52abdf7bb7e01392024-12-19T11:01:49ZengElsevierPlant Stress2667-064X2024-12-0114100575Identification of reference genes via real-time quantitative PCR for investigation of the transcriptomic basis of the squalene biosynthesis in different tissues on olives under drought stressHaiyang Chi0Shasha Bai1Chenxiao Chen2Yuying Bao3Xiaoxin Qu4Shitao Sun5Jiangpeng Pan6Xiushi Yang7Chunsheng Hou8Yanchun Deng9Inner Mongolia University, College of Life Sciences, Hohhot 010000, PR China; Chinese Academy of Agricultural Sciences, Center of Southern Economic Crops, Changsha 410205, PR China; Chinese Academy of Agricultural Sciences, Institute Bast Fiber Crops, Changsha 410205, PR ChinaBeijing Institute of Nutritional Resources Co., LTD, Beijing Academy of Science and Technology, Beijing 100069, PR ChinaChinese Academy of Agricultural Sciences, Institute Bast Fiber Crops, Changsha 410205, PR ChinaInner Mongolia University, College of Life Sciences, Hohhot 010000, PR ChinaChinese Academy of Agricultural Sciences, Institute Bast Fiber Crops, Changsha 410205, PR China; Chinese Academy of Agricultural Sciences, Center of Southern Economic Crops, Changsha 410205, PR ChinaChinese Academy of Agricultural Sciences, Institute Bast Fiber Crops, Changsha 410205, PR China; Chinese Academy of Agricultural Sciences, Center of Southern Economic Crops, Changsha 410205, PR ChinaChinese Academy of Agricultural Sciences, Institute Bast Fiber Crops, Changsha 410205, PR China; Chinese Academy of Agricultural Sciences, Center of Southern Economic Crops, Changsha 410205, PR ChinaChinese Academy of Agricultural Sciences, Institute Bast Fiber Crops, Changsha 410205, PR China; Chinese Academy of Agricultural Sciences, Center of Southern Economic Crops, Changsha 410205, PR ChinaChinese Academy of Agricultural Sciences, Institute Bast Fiber Crops, Changsha 410205, PR China; Chinese Academy of Agricultural Sciences, Center of Southern Economic Crops, Changsha 410205, PR China; Corresponding authors.Chinese Academy of Agricultural Sciences, Institute Bast Fiber Crops, Changsha 410205, PR China; Chinese Academy of Agricultural Sciences, Center of Southern Economic Crops, Changsha 410205, PR China; Corresponding authors.Squalene plays a crucial role in plant growth, development, and stress tolerance. To select the best reference genes (RGs) for expression profile analysis of genes involved in squalene biosynthesis in olives under stress tolerance, the expression stability of 22 candidate RGs across four tissues (root, stem, leaf, and fruit) and five representative cultivars under drought stress was assessed using six methods. Our study showed that, ubiquitin-conjugating enzyme1 (UBC1) and 60S ribosomal protein L18-2-like (60S) were the most stable RGs across five different cultivars and various tissues. Additionally, elongation factor 1-alpha-like (EF1-α2) and UBC1 were the most stable RG in olives under drought stress. UBC1 was the appropriate RG for further study, qPCR analysis showed that five certain genes involved in the squalene biosynthesis exhibited significant differential expression under varying conditions. Specially, the expression level of squalene synthase (SQS) in the leaves was highest, and HPLC analysis showed that squalene content was the highest in leaves. Likewise, the expression levels of SQS and farnesyl diphosphate synthase (FPPS) in the leaves of the cultivar Arbosana and Chelmsford Lal were significantly higher than those of the other cultivars, respectively, and HPLC analysis also showed that squalene content was highest in the two cultivars. Interestingly, the amount of squalene increased dramatically in olive under drought stress, as lupeol synthase (LUPS) was significantly up-regulated. To date, this study firstly provided the comprehensive analysis of RGs related to squalene biosynthesis in various cultivars and tissues of olives under drought stress.http://www.sciencedirect.com/science/article/pii/S2667064X24002288Reference genesOlivesGenes expressionSqualene biosynthesisDrought stress
spellingShingle Haiyang Chi
Shasha Bai
Chenxiao Chen
Yuying Bao
Xiaoxin Qu
Shitao Sun
Jiangpeng Pan
Xiushi Yang
Chunsheng Hou
Yanchun Deng
Identification of reference genes via real-time quantitative PCR for investigation of the transcriptomic basis of the squalene biosynthesis in different tissues on olives under drought stress
Plant Stress
Reference genes
Olives
Genes expression
Squalene biosynthesis
Drought stress
title Identification of reference genes via real-time quantitative PCR for investigation of the transcriptomic basis of the squalene biosynthesis in different tissues on olives under drought stress
title_full Identification of reference genes via real-time quantitative PCR for investigation of the transcriptomic basis of the squalene biosynthesis in different tissues on olives under drought stress
title_fullStr Identification of reference genes via real-time quantitative PCR for investigation of the transcriptomic basis of the squalene biosynthesis in different tissues on olives under drought stress
title_full_unstemmed Identification of reference genes via real-time quantitative PCR for investigation of the transcriptomic basis of the squalene biosynthesis in different tissues on olives under drought stress
title_short Identification of reference genes via real-time quantitative PCR for investigation of the transcriptomic basis of the squalene biosynthesis in different tissues on olives under drought stress
title_sort identification of reference genes via real time quantitative pcr for investigation of the transcriptomic basis of the squalene biosynthesis in different tissues on olives under drought stress
topic Reference genes
Olives
Genes expression
Squalene biosynthesis
Drought stress
url http://www.sciencedirect.com/science/article/pii/S2667064X24002288
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