An Analysis of Alternative Splicing Events During Browning Inhibition of Fresh-cut Apples by Hydrogen Sulfide Treatment

doi:10.16420/j.issn.0513-353x.2020-0847

Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (11): 2121-2132.doi: 10.16420/j.issn.0513-353x.2020-0847

• Original article • Next Articles

An Analysis of Alternative Splicing Events During Browning Inhibition of Fresh-cut Apples by Hydrogen Sulfide Treatment

CHEN Chen, LIU Chenghui, SHI Lijia, JIANG Aili(), HU Wenzhong()

Key Laboratory of Biotechnology and Bioresources Utilization,Ministry of Education,College of Life Science,Dalian Minzu University,Dalian,Liaoning 116600,China

Received:2021-06-11 Revised:2021-08-06 Published:2021-12-02
Contact: JIANG Aili,HU Wenzhong E-mail:jal@dlnu.edu.cn;hwz@dlnu.edu.cn

Abstract

Abstract:

In order to explore the alternative splicing of the browning inhibition of fresh-cut apples by hydrogen sulfide（H₂S）treatment,RNA-Seq technology was used to identify the alternative splicing occurring in H₂S-treated and H₂O-treated fresh-cut apples at two storage time（0 and 6 d）. The results showed that the dominate alternative splicing types were alternative transcription start site（TSS）and alternative transcription termination site（TTS）in these four samples. The number of the specific alternative splicing genes of H₂S-treated fresh-cut apples at 0 and 6 d were 921 and 2 423,respectively. GO annotation showed that the specific alternative splicing genes in H₂S-treated fresh-cut apples at 0 and 6 d were mainly enriched in flavonoid biosynthetic process,cytoplasm,membrane,hydrolase and oxidoreductase. KEGG analysis showed that these genes mainly enriched in plant-pathogen interaction,plant hormone signal transduction,carbohydrate metabolism,phenylpropanoid biosynthesis and oxidative phosphorylation. Furthermore,expression of polyphenol oxidase（PPO）,peroxidase（POD）,lipoxidase （LOX）and phospholipase D（LPD）genes was down-regulated after H₂S treatment,whilst expression of phenylalanine ammonia-lyase（PAL）and NADH dehydrogenase was induced. In conclusion,H₂S controlled the browning of fresh-cut apples through changing the alternative splicing and expression pattern of peroxide,phenylpropanoid metabolism,membrane and energy metabolism related genes.

Key words: apple, fresh-cut, browning, hydrogen sulfide, alternative splicing

CLC Number:

S661.1

CHEN Chen, LIU Chenghui, SHI Lijia, JIANG Aili, HU Wenzhong. An Analysis of Alternative Splicing Events During Browning Inhibition of Fresh-cut Apples by Hydrogen Sulfide Treatment[J]. Acta Horticulturae Sinica, 2021, 48(11): 2121-2132.

Figures/Tables 8

References 28

[1]	Hu Wenzhong, Jiang Aili, Liu Chenghui, Zhao Lei. 2018. Physiological mechanism for browning inhibition in fresh-cut apple by cysteine. Food Science, 39 (3):282-288. (in Chinese) doi: 10.1111/jfds.1974.39.issue-2 URL
	陈晨, 胡文忠, 姜爱丽, 刘程惠, 赵蕾. 2018. 半胱氨酸控制鲜切苹果褐变的生理机制. 食品科学, 39 (3):282-288.
[2]	Cortellino G, Gobbi S, Bianchi G, Rizzolo A. 2015. Modified atmosphere packaging for shelf life extension of fresh-cut apples. Trends in Food Science and Technology, 46 (2):320-330. doi: 10.1016/j.tifs.2015.06.002 URL
[3]	Docimo T, Francese G, de Palma M, Mennella D, Toppion L, Scalzo P L, Mennella G, Tucci M. 2016. Insights in the fruit flesh browning mechanisms in Solanum melongena genetic lines with opposite postcut behavior. Journal of Agricultural and Food Chemistry, 64 (22):4675-4685. doi: 10.1021/acs.jafc.6b00662 pmid: 27198496
[4]	Feng Yalan, Xiong Ying, Zhang Jun, Yuan Jiale, Cai Aishan, Ma Chao. 2020. Role of alternative splicing in plant development and abiotic stress responses. Journal of Nuclear Agricultural Sciences, 34 (1):62-70. (in Chinese)
	冯雅岚, 熊瑛, 张均, 原佳乐, 蔡艾杉, 马超. 2020. 可变剪切在植物发育和非生物胁迫响应中的作用. 核农学报, 34 (1):62-70.
[5]	Franck C, Lammertyn J, Ho Q T, Verboven P, Verlinden B, Nicolai B M. 2007. Browning disorders in pear fruit. Postharvest Biology and Technology, 43 (1):1-13. doi: 10.1016/j.postharvbio.2006.08.008 URL
[6]	Hancock J T, Whiteman M. 2014. Hydrogen sulfide and cell signaling: team player or referee? Plant Physiology and Biochemistry, 78:37-42.
[7]	Hu H, Shen W, Li P. 2014. Effects of hydrogen sulphide on quality and antioxidant capacity of mulberry fruit. International Journal of Food Science and Technology, 49:399-409. doi: 10.1111/ijfs.12313 URL
[8]	Hu L, Hu S L, Wu J, Li Y H, Zheng J L, Wei Z J, Liu J, Wang H L, Liu Y S, Zhang H. 2012. Hydrogen sulfide prolongs postharvest shelf life of strawberry and plays an antioxidative role in fruits. Journal of Agriculture and Food Chemistry, 60:8684-8693. doi: 10.1021/jf300728h URL
[9]	Huang Ling, Li Qiongying, Wei Shugu, Lai Jia, Dai Shundong, Zhang Qianfang, Zeng Hualan, Liu Jia, Ye Pengsheng. 2019. Identification and difference analysis of the alternative splicing event in the hermaphroditic flowers and male flowers of Asparagus officinalis. Acta Horticulturae Sinica, 46 (8):1503-1518. (in Chinese)
	黄玲, 李琼英, 韦树谷, 赖佳, 代顺冬, 张骞方, 曾华兰, 刘佳, 叶鹏盛. 2019. 芦笋两性花与雄花中发生可变剪接基因的差异分析. 园艺学报, 46 (8):1503-1518.
[10]	Jiang J, Jiang L, Zhang L, Luo H, Opiyo A M, Yu Z. 2012. Changes of protein profile in fresh-cut lotus tuber before and after browning. Journal of Agricultural and Food Chemistry, 60 (15):3955-3965. doi: 10.1021/jf205303y pmid: 22455495
[11]	Jin Z, Pei Y. 2015. Physiological implications of hydrogen sulfide in plants: pleasant exploration behind its unpleasant odour. Oxidative Medicine and Cellular Longevity, 2015:397502.
[12]	Laloum T, Martín G, Duque P. 2018. Alternative splicing control of abiotic stress responses. Trends in Plant Science, 23:140-150. doi: S1360-1385(17)30218-2 pmid: 29074233
[13]	Landrigan M, Morris S C, Eamus D, McGlasson W B. 1996. Postharvest water relationships and tissue browning of rambutan fruit. Scientia Horticulturae, 66 (3-4):201-208. doi: 10.1016/S0304-4238(96)00915-6 URL
[14]	Liang D, Wang C, Tocmo R, Wu H, Deng L W, Huang D. 2015. Hydrogen sulphide(H₂S)releasing capacity of essential oils isolated from organosulphur rich fruits and vegetables. Journal of Functional Foods, 14:634-640. doi: 10.1016/j.jff.2015.02.007 URL
[15]	Lin Y, Lin Y, Lin H, Chen Y, Wang H, Shi J. 2018. Application of propyl gallate alleviates pericarp browning in harvested longan fruit by modulating metabolisms of respiration and energy. Food Chemistry, 240:863-869. doi: 10.1016/j.foodchem.2017.07.118 URL
[16]	Mellidou I, Buts K, Hatoum D, Ho Q T, Johnston J W, Watkins C B, Schaffer R J, Gapper N E, Giovannoni J J, Rudell D R, Hertog M L, Nicolai B M. 2014. Transcriptomic events associated with internal browning of apple during postharvest storage. BMC Plant Biology, 14:328. doi: 10.1186/s12870-014-0328-x URL
[17]	Milani J, Hamedi M. 2004. Susceptibility of five apple cultivars to enzymatic browning. International Postharvest Symposium,2221-2226.
[18]	Qiao D, Yang C, Chen J, Guo Y, Li Y, Niu S, Cao K, Chen Z. 2019. Comprehensive identification of the full-length transcripts and alternative splicing related to the secondary metabolism pathways in the tea plant(Camellia sinensis). Scientific Reports, 9 (1):1-13.
[19]	Rawyler A, Pavelic D, Gianinazzi C, Oberson J, Braendle R. 1999. Membrane lipid integrity relies on a threshold of ATP production rate in potato cell cultures submitted to anoxia. Plant Physiology, 120 (1):93-300. doi: 10.1104/pp.120.1.93 URL
[20]	Reddy A S N, Marquez Y, Kalyna M, Barta A. 2013. Complexity of the alternative splicing landscape in plants. The Plant Cell, 25 (10):3657-3683. doi: 10.1105/tpc.113.117523 URL
[21]	Saquet A A, Streif J, Bangerth F. 2003. Energy metabolism and membrane lipid alterations in relation to brown heart development in‘Conference’pears during delayed controlled atmosphere storage. Postharvest Biology and Technology, 30 (2):123-132. doi: 10.1016/S0925-5214(03)00099-1 URL
[22]	Sun Y, Zhang W, Zeng T, Nie Q, Zhang F, Zhu L. 2015. Hydrogen sulfide inhibits enzymatic browning of fresh-cut lotus root slices by regulating phenolic metabolism. Food Chemistry, 177:376-381. doi: 10.1016/j.foodchem.2015.01.065 URL
[23]	Tan Yitan, Zeng Kaifang. 2014. Effects of combined treatment with ascorbic acid,cysteine and CaCl₂ on browning of fresh-cut taro. Food Science, 35 (4):231-235. (in Chinese)
	谭谊谈, 曾凯芳. 2014. 抗坏血酸、半胱氨酸与氯化钙复合处理对鲜切芋艿褐变的影响. 食品科学, 35 (4):231-235.
[24]	Toivonen P M A, Brummell D A. 2008. Biochemical bases of appearance and texture changes in fresh-cut fruit and vegetables. Postharvest Biology and Technology, 48 (1):1-14. doi: 10.1016/j.postharvbio.2007.09.004 URL
[25]	Wang R. 2002. Two’s company,three’sa crowd: can H₂S be the third endogenous gaseous transmitter? The FASEB Journal, 16:1792-1798. doi: 10.1096/fsb2.v16.13 URL
[26]	Zhang Xiaoyan, Liu Aiwen, Ji Qiyan, Peng Yong. 2020. Effect of heat treatment combined with γ-aminobutyric acid on the physiology property and quality of fresh-cut apples. Science and Technology of Food Industry, 41 (14):265-269,275. (in Chinese)
	张小燕, 刘艾雯, 籍奇岩, 彭勇. 2020. 热处理结合γ-氨基丁酸对鲜切苹果生理特性和品质的影响. 食品工业科技, 41 (14):265-269,275.
[27]	Zhang J L, Hu L Y, Hu K D, Wu J, Yang F, Zhang H. 2016. Hydrogen sulfide alleviates senescence of fresh-cut apple by regulating antioxidant defense system and senescence-related gene expression. HortScience, 51:152-158. doi: 10.21273/HORTSCI.51.2.152 URL
[28]	Zhou Qi, Zhou Fuhui, Hu Wenzhong, Zhao Lei, Xu Yuanyuan. 2019. Correlation between enzymatic browning inhibition by UV-C treatment and reactive oxygen species metabolism of fresh-cut apples. Food Science, 40 (5):110-117. (in Chinese)
	周琪, 陈晨, 周福慧, 胡文忠, 赵蕾, 许源源. 2019. 短波紫外线控制鲜切苹果褐变与其活性氧代谢的相关性. 食品科学, 40 (5):110-117.

基因 Gene	上游引物（5′-3′） Forward primer	下游引物（5′-3′） Reverse primer
PPO	GTGGCTTGCGGTATTATTGGT	GATGCTCGGTTTCCCTCAA
POD	CCAACCTCGCAACCCTAATCT	CACTGAGACTGGCCTAATGTGTG
PAL	CGCTGGAGTGTTTGGAAGTG	AGTAGCCTTGGAGGAGTGTGTTG
PLD	GTCTTGATCTTTGTGATGGTCGTT	GGTTGTCTTGGAGCCTTGGT
LOX	AGCTTGGGCTGAAAATCCTGT	ACAACGCCTGCTCCAACTCT
NADH脱氢酶基因NADH dehydrogenase	TGGTATCGGACTGGGGTAGG	ATGTGCGTCCAAGTGTTTGTG
18s RNA	GATTCCGGTGCCCAGAAGT	CCAGCAGCTTCCATTCCAA

基因 Gene	上游引物（5′-3′） Forward primer	下游引物（5′-3′） Reverse primer
PPO	GTGGCTTGCGGTATTATTGGT	GATGCTCGGTTTCCCTCAA
POD	CCAACCTCGCAACCCTAATCT	CACTGAGACTGGCCTAATGTGTG
PAL	CGCTGGAGTGTTTGGAAGTG	AGTAGCCTTGGAGGAGTGTGTTG
PLD	GTCTTGATCTTTGTGATGGTCGTT	GGTTGTCTTGGAGCCTTGGT
LOX	AGCTTGGGCTGAAAATCCTGT	ACAACGCCTGCTCCAACTCT
NADH脱氢酶基因NADH dehydrogenase	TGGTATCGGACTGGGGTAGG	ATGTGCGTCCAAGTGTTTGTG
18s RNA	GATTCCGGTGCCCAGAAGT	CCAGCAGCTTCCATTCCAA

样品Sample			Raw reads 原始数据	Clean reads 有效数据	质量值 ≥ 20的碱基所占比例/% Q20	质量值 ≥ 30的碱基所占比例/% Q30	GC含量所占的比例/% GC content
处理 Treatment		天数/d Day	Raw reads 原始数据	Clean reads 有效数据	质量值 ≥ 20的碱基所占比例/% Q20	质量值 ≥ 30的碱基所占比例/% Q30	GC含量所占的比例/% GC content
H₂O	0		60 422 744	59 865 744	99.61	95.03	47.50
			51 249 354	50 902 542	99.75	95.56	47.00
			44 214 706	43 903 682	99.73	95.47	47.00
	6		44 706 736	44 404 318	99.66	95.07	47.00
			40 462 228	40 210 134	99.66	95.04	47.00
			47 660 554	47 291 638	99.70	95.33	47.00
H₂S	0		45 000 228	44 602 378	99.71	95.17	47.00
			50 155 628	49 585 722	99.74	95.94	47.00
			45 750 610	45 333 712	99.73	95.78	47.00
	6		47 041 536	46 731 660	99.67	95.17	47.00
			46 388 294	46 024 028	99.76	95.62	47.00
			52 755 840	52 352 240	99.43	96.58	47.00

样品Sample			Raw reads 原始数据	Clean reads 有效数据	质量值 ≥ 20的碱基所占比例/% Q20	质量值 ≥ 30的碱基所占比例/% Q30	GC含量所占的比例/% GC content
处理 Treatment		天数/d Day	Raw reads 原始数据	Clean reads 有效数据	质量值 ≥ 20的碱基所占比例/% Q20	质量值 ≥ 30的碱基所占比例/% Q30	GC含量所占的比例/% GC content
H₂O	0		60 422 744	59 865 744	99.61	95.03	47.50
			51 249 354	50 902 542	99.75	95.56	47.00
			44 214 706	43 903 682	99.73	95.47	47.00
	6		44 706 736	44 404 318	99.66	95.07	47.00
			40 462 228	40 210 134	99.66	95.04	47.00
			47 660 554	47 291 638	99.70	95.33	47.00
H₂S	0		45 000 228	44 602 378	99.71	95.17	47.00
			50 155 628	49 585 722	99.74	95.94	47.00
			45 750 610	45 333 712	99.73	95.78	47.00
	6		47 041 536	46 731 660	99.67	95.17	47.00
			46 388 294	46 024 028	99.76	95.62	47.00
			52 755 840	52 352 240	99.43	96.58	47.00

An Analysis of Alternative Splicing Events During Browning Inhibition of Fresh-cut Apples by Hydrogen Sulfide Treatment

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