Transcriptome Analysis of the Molecular Mechanism of Saline-alkali Stress Response in Malus halliana Leaves

doi:10.16420/j.issn.0513-353x.2021-0077

Acta Horticulturae Sinica ›› 2022, Vol. 49 ›› Issue (2): 237-251.doi: 10.16420/j.issn.0513-353x.2021-0077

• Research Papers • Next Articles

Transcriptome Analysis of the Molecular Mechanism of Saline-alkali Stress Response in Malus halliana Leaves

ZHANG Rui, ZHANG Xiayi, ZHAO Ting, WANG Shuangcheng, ZHANG Zhongxing, LIU Bo, ZHANG De, WANG Yanxiu()

College of Horticulture,Gansu Agricultural University,Lanzhou 730070,China

Received:2021-05-11 Revised:2021-07-19 Online:2022-02-25 Published:2022-02-28
Contact: WANG Yanxiu E-mail:wangxy@gsau.edu.cn

Abstract

Abstract:

Malus halliana is a highly saline-alkali-resistant apple rootstock in north-western China. The genes and main pathways which respond to saline-alkali stress were identified by RNA-Seq. This study revealed the mechanism of the response to salt damage and provided theoretical basis for the salt tolerance mechanism of apple rootstocks. Normal water（Hoagland nutrient solution）（Control）supply and mixed saline-alkali stress（Hoagland nutrient solution + 100 mmol · L^-1 NaCl + NaHCO₃）（Treatment）were set up to screen the saline-alkali resistance of each treatment in the growth period. These two samples before and after saline-alkali treatment were analyzed by RNA-seq. The function and pathway enrichment of differentially expressed genes（DEG）were also performed by GO and KEGG analysis. Twenty DEG were randomly selected for further qRT-PCR analysis to verify the RNA-seq data. A total of 16 246 DEGs were determined,7 268 genes were up-regulated and 8 978 genes down-regulated under saline-alkali stress. GO analysis fund that the DEGs were considerably differenct in the biological processes,cell components and molecular functions. KEGG analysis indicated that most enriched saline-alkali-responsive genes were mainly involved in signal transduction,carbon metabolism,biosynthesis of amino acids,and other secondary metabolites. DEG mainly focused on calcium signaling pathway,plant hormone signal transduction,phenylalanine metabolism,and carotenoid biosynthesis. qRT-PCR detection results of 20 DEG expression patterns were similar to those of RNA-seq. Among them,aspartate aminotransferase（GOT1）,monodehydroascorbate reductase（NADH）,chalcone isomerase（CHI,E5.5.1.6）,β-carotene 3-hydroxylase（CrtZ）genes were highly expressed during saline-alkali stress. The results indicated that they might play a role in the leaf response to saline-alkali stress. M. halliana is mainly regulated by calcium signal pathway,plant hormone signal transduction,biosynthesis of amino acids, carotenoid biosynthesis and other secondary metabolites to respond to saline-alkali stress.

Key words: Malus halliana, saline-alkali stress, transcriptomic profiles analysis, differential gene expression

CLC Number:

S661.4

ZHANG Rui, ZHANG Xiayi, ZHAO Ting, WANG Shuangcheng, ZHANG Zhongxing, LIU Bo, ZHANG De, WANG Yanxiu. Transcriptome Analysis of the Molecular Mechanism of Saline-alkali Stress Response in Malus halliana Leaves[J]. Acta Horticulturae Sinica, 2022, 49(2): 237-251.

Figures/Tables 9

References 41

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基因 Gene	名称 Description	上游引物（5′-3′） Forward primer	下游引物（5′-3′） Reverse primer
ANT	溶质载体家族25 Solute carrier family 25	CCAACTTCCACCGTGATCTGATG	TCCTTCCACCGCCGAGAATG
ATP2A	钙转运体 Calcium transporter	GGTACAGTAGTCGTGGCTGGTAG	TTCAATGGCGTCACTTCATCTTCC
PLCD	磷脂酰肌醇磷脂酶C Phosphatidylinositol phospholipase C,delta	GGTGTTCGCCAAGTACGCTCAG	GGGTCGTGAACTGGAGGAGGATAG
SPHK	鞘氨醇激酶Sphingosine kinase	CTCATAAGCGGCGTCGTCACAC	CGAACCCCAGGACCTCCTTCTC
VDAC1	赖性阴离子通道蛋白1 Voltage-dependent anion channel protein 1	TGCCGAACTGACCCACAGATTTTC	GGTCTCCATTCACGCTGCCATAG
ABF	ABA响应元件结合因子 ABA responsive element binding factor	TGACAATGTTGAGTTCCGTGAGC	TCGACGACTGCCTCGCTAAC
PR1	病程相关蛋白1 Pathogenesis-related protein 1	ATGGGGTTGTGCAATATTTC	CTAGTAAGGCTTCTCCCCAAC
PYL	脱落酸受体PYR/PYL家族 Abscisic acid receptor PYR/PYL family	ATGTCTTCACCAATCCAGTTTC	AGTACTCCCAAAGAACCACAAA
COI-1	冠菌素不敏感蛋白1 Coronatine-insensitive protein 1	CGAGGGCGGCGATGTTCAATC	ACTTCAAGCGGTGGAACGAGTTG
GOT1	细胞质天冬氨酸转氨酶 Aspartate aminotransferase,cytoplasmic	ACAACGCCTCCAAGCTCCTC	TCTGAGTTCGCCGAGTCCTTC
CHI	查耳酮异构酶Chalcone isomerase	ATGGCTGACGGTGACCAAC	TTATCTAGTGATATGAGCAATAAA
ANS	花青素合成酶Anthocyanidin synthase	ATGGTGAGCTCTGATTCAG	TCACTTGGGGAGCAAAGC
ANR	花青素还原酶Anthocyanidin reductase	ATGGCCACCCAACAACCC	CTAGTTCTGCAGCAGCCC
NADH	单脱水抗坏血酸还原酶 Monodehydroascorbate reductase（NADH）	ATGGGAAGGGCATTTGTG	TCACCACCTCCGGCGCTT
CrtY	番茄红素Lycopene beta-cyclase	ATGGATACATTGCTTAAAACGC	GATTAGAGATAGGACATAGTTCAAC
CrtZ	β-胡萝卜素β-carotene 3-hydroxylase	GCAAGGTGGGCTCATAGAACTCTG	AAGAAGGGCAATAGCAGGAACAGC
HPD	羟苯丙酮酸加双氧酶 Hydroxyphenylpyruvate dioxygenase	TGTACCACAGAGACCTGAAGCC	CAGCAGCAGAGCCATCATAACC
pckA	膜伯胺氧化酶 Primary-amine oxidase	CGACGCCTTCCTGTTGCTTCC	CGGTGACGCCAGTCCAATCTTG
VPS35	液泡蛋白分选相关蛋白35 Vacuolar protein sorting-associated protein 35	TGAAGATTGGCTGCACTTGACTGG	CCGACAACTGCCCATTGAGGAAG
VTA1	液泡蛋白分选相关蛋白 Vacuolar protein sorting-associated protein	GGGCTCTGATCCTTCCTACTCTCC	AGTCCCATTTCTGTTGCTCGTCTG

基因 Gene	名称 Description	上游引物（5′-3′） Forward primer	下游引物（5′-3′） Reverse primer
ANT	溶质载体家族25 Solute carrier family 25	CCAACTTCCACCGTGATCTGATG	TCCTTCCACCGCCGAGAATG
ATP2A	钙转运体 Calcium transporter	GGTACAGTAGTCGTGGCTGGTAG	TTCAATGGCGTCACTTCATCTTCC
PLCD	磷脂酰肌醇磷脂酶C Phosphatidylinositol phospholipase C,delta	GGTGTTCGCCAAGTACGCTCAG	GGGTCGTGAACTGGAGGAGGATAG
SPHK	鞘氨醇激酶Sphingosine kinase	CTCATAAGCGGCGTCGTCACAC	CGAACCCCAGGACCTCCTTCTC
VDAC1	赖性阴离子通道蛋白1 Voltage-dependent anion channel protein 1	TGCCGAACTGACCCACAGATTTTC	GGTCTCCATTCACGCTGCCATAG
ABF	ABA响应元件结合因子 ABA responsive element binding factor	TGACAATGTTGAGTTCCGTGAGC	TCGACGACTGCCTCGCTAAC
PR1	病程相关蛋白1 Pathogenesis-related protein 1	ATGGGGTTGTGCAATATTTC	CTAGTAAGGCTTCTCCCCAAC
PYL	脱落酸受体PYR/PYL家族 Abscisic acid receptor PYR/PYL family	ATGTCTTCACCAATCCAGTTTC	AGTACTCCCAAAGAACCACAAA
COI-1	冠菌素不敏感蛋白1 Coronatine-insensitive protein 1	CGAGGGCGGCGATGTTCAATC	ACTTCAAGCGGTGGAACGAGTTG
GOT1	细胞质天冬氨酸转氨酶 Aspartate aminotransferase,cytoplasmic	ACAACGCCTCCAAGCTCCTC	TCTGAGTTCGCCGAGTCCTTC
CHI	查耳酮异构酶Chalcone isomerase	ATGGCTGACGGTGACCAAC	TTATCTAGTGATATGAGCAATAAA
ANS	花青素合成酶Anthocyanidin synthase	ATGGTGAGCTCTGATTCAG	TCACTTGGGGAGCAAAGC
ANR	花青素还原酶Anthocyanidin reductase	ATGGCCACCCAACAACCC	CTAGTTCTGCAGCAGCCC
NADH	单脱水抗坏血酸还原酶 Monodehydroascorbate reductase（NADH）	ATGGGAAGGGCATTTGTG	TCACCACCTCCGGCGCTT
CrtY	番茄红素Lycopene beta-cyclase	ATGGATACATTGCTTAAAACGC	GATTAGAGATAGGACATAGTTCAAC
CrtZ	β-胡萝卜素β-carotene 3-hydroxylase	GCAAGGTGGGCTCATAGAACTCTG	AAGAAGGGCAATAGCAGGAACAGC
HPD	羟苯丙酮酸加双氧酶 Hydroxyphenylpyruvate dioxygenase	TGTACCACAGAGACCTGAAGCC	CAGCAGCAGAGCCATCATAACC
pckA	膜伯胺氧化酶 Primary-amine oxidase	CGACGCCTTCCTGTTGCTTCC	CGGTGACGCCAGTCCAATCTTG
VPS35	液泡蛋白分选相关蛋白35 Vacuolar protein sorting-associated protein 35	TGAAGATTGGCTGCACTTGACTGG	CCGACAACTGCCCATTGAGGAAG
VTA1	液泡蛋白分选相关蛋白 Vacuolar protein sorting-associated protein	GGGCTCTGATCCTTCCTACTCTCC	AGTCCCATTTCTGTTGCTCGTCTG

样品 Sample	原始数据 Raw reads	过滤数据 Clean reads	有效碱基/% Valid bases	Q30含量/% Q30 content	GC含量/% GC content
对照 Control	49 794 146	47 025 046	90.15	93.52	47.70
胁迫处理 Stress treatment	49 258 400	47 513 110	92.50	94.63	47.90

样品 Sample	原始数据 Raw reads	过滤数据 Clean reads	有效碱基/% Valid bases	Q30含量/% Q30 content	GC含量/% GC content
对照 Control	49 794 146	47 025 046	90.15	93.52	47.70
胁迫处理 Stress treatment	49 258 400	47 513 110	92.50	94.63	47.90

Transcriptome Analysis of the Molecular Mechanism of Saline-alkali Stress Response in Malus halliana Leaves

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