NaCl处理下全缘冬青和红果冬青根系的转录组活性比较

doi:10.12302/j.issn.1000-2006.202109054

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南京林业大学学报（自然科学版） ›› 2022, Vol. 46 ›› Issue (3): 99-108.doi: 10.12302/j.issn.1000-2006.202109054

NaCl处理下全缘冬青和红果冬青根系的转录组活性比较

张强¹(), 周鹏(), 刘昌来¹, 余永帆¹, 张敏²^,^*(), 杨甲定¹^,^*()

1.南京林业大学,南方现代林业协同创新中心,南京林业大学生物与环境学院,江苏南京 210037
2.江苏省林业科学研究院,江苏南京 211153

收稿日期:2021-09-27 接受日期:2021-11-08 出版日期:2022-05-30 发布日期:2022-06-10
通讯作者: 张敏,杨甲定
基金资助:
江苏省林业科技创新与推广项目(LYKJ[2021]07);江苏省林业科学研究院自主科研项目(ZZKY202105)

Comparison of transcriptomic activity of Ilex integra and I. purpurea roots with NaCl treatments

ZHANG Qiang¹(), ZHOU Peng(), LIU Changlai¹, YU Yongfan¹, ZHANG Min²^,^*(), YANG Jiading¹^,^*()

1. Co-Innovation Center for Sustainable Forestry in Southern China, College of Biology and the Environment, Nanjing Forestry University, Nanjing 210037, China
2. Jiangsu Academy of Forestry, Nanjing 211153, China

Received:2021-09-27 Accepted:2021-11-08 Online:2022-05-30 Published:2022-06-10
Contact: ZHANG Min,YANG Jiading

摘要/Abstract

摘要：

【目的】研究全缘冬青(Ilex integra)和红果冬青(I. purpurea)的根系在NaCl胁迫处理中的转录组活性差异,为进一步研究二者耐盐性差异的生理学机理奠定基础。【方法】以内含250 mmol/L NaCl的1/2浓度霍格兰德溶液对全缘冬青和红果冬青的两年生扦插苗进行盐胁迫处理,采集经0(盐处理前)、6和72 h盐处理的根系样品,提取总RNA,进行转录组测序,组装转录本,筛选差异表达基因,分析两种冬青根系中的功能富集通路和特定代谢途径的基因表达变化。【结果】全缘冬青根系盐处理6 h的差异表达基因数量为2 616,盐处理72 h为1 802;而红果冬青根系盐处理6 h的差异表达基因数量为1 831,盐处理72 h为3 490。全缘冬青盐处理6和72 h根系中共同的KEGG富集途径为植物激素信号转导、亚油酸代谢、类胡萝卜素生物合成和甜菜红素生物合成,红果冬青根系6和72 h盐处理共同的KEGG富集途径为植物激素信号转导、苯丙烷生物合成和类胡萝卜素生物合成。甜菜红素生物合成和亚油酸/花生四烯酸代谢为盐胁迫处理全缘冬青根系中特有的富集代谢通路。全缘冬青中受盐胁迫正调控的过氧化氢酶基因数多于红果冬青的,并且在盐处理下其表达水平远高于红果冬青的。【结论】全缘冬青根系较强的耐盐性可能与植物激素信号转导、类胡萝卜素生物合成、甜菜红素生物合成和脂肪酸脂类代谢等生理过程相关联;盐处理下过氧化氢酶基因的高效表达也可能是全缘冬青根系具较强耐盐性的因素之一。

关键词: 全缘冬青, 红果冬青, 根系, NaCl盐胁迫, 转录组分析, 代谢途径

Abstract:

【Objective】 This research aims to investigate the difference in transcriptomic activity of Ilex integra and I. purpurea roots using NaCl salt treatments and provide insights for future study on the physiological mechanisms responsible for differential salt tolerance between these Ilex species. 【Method】 Two-year-old cutting seedlings of I. integra and I. purpurea were irrigated with half-strength Hoagland solution containing 250 mmol/L NaCl for salt treatments. Root samples were collected at 0(before salt treatment), 6 and 72 h after salt treatments. The total RNAs were extracted for transcriptomic RNA-Seq. Transcripts were assembled and differentially expressed genes (DEGs) were selected. The enriched KEGG functional pathways for the two species were compared, and the expression of genes involved in certain metabolic pathways was analyzed. 【Result】 There were 2 616 and 1 802 DEGs in 6 h-salt-treated and 72 h-salt-treated I. integra roots respectively, while there were 1 831 and 3 490 DEGs in 6 h-salt-treated and 72 h-salt-treated I. purpurea roots, respectively. The common enriched KEGG functional pathways between I. integra roots with 6 and 72 h salt treatments were plant hormone signal transduction, linoleic acid metabolism, carotenoid biosynthesis and betalain biosynthesis. The common enriched pathways between I. integra roots with 6 and 72 h salt treatments were plant hormone signal transduction, phenylpropanoid biosynthesis and carotenoid biosynthesis. Betalain biosynthesis and linoleic acid/arachidonic acid metabolism are unique pathways in I. integra roots during salt treatments. There were more up-regulated putative catalase genes with much higher expression levels in salt-treated I. integra roots than that in the I. purpurea roots. 【Conclusion】 The higher salt tolerance of I. integra roots may be associated with physiological processes such as plant hormone signal transduction, carotenoid biosynthesis, betalain biosynthesis and fatty acid metabolism. The higher expression of catalase genes during salt treatments may also contribute to the higher salt tolerance of I. integra roots.

Key words: Ilex integra, I. purpurea, roots, NaCl salt treatment, transcriptomic analysis, metabolic pathway

中图分类号:

Q945
S718

张强,周鹏,刘昌来,等. NaCl处理下全缘冬青和红果冬青根系的转录组活性比较[J]. 南京林业大学学报（自然科学版）, 2022, 46(3): 99-108.

ZHANG Qiang, ZHOU Peng, LIU Changlai, YU Yongfan, ZHANG Min, YANG Jiading. Comparison of transcriptomic activity of Ilex integra and I. purpurea roots with NaCl treatments[J].Journal of Nanjing Forestry University (Natural Science Edition), 2022, 46(3): 99-108.DOI: 10.12302/j.issn.1000-2006.202109054.

图/表 7

表1

荧光定量PCR 引物"

类别 type	基因 gene	基因号 gene ID		正向引物 forward primer (5'—3')	反向引物 reverse primer (5'—3')
全缘冬青甜菜红素合成相关基因 betalain biosynthesis-related genes in I. integra	Tyrosinase-like	TRINITY_DN62285_c0_g3		ACATGGCTTTCCCTCCTGTT	AGAATGGGTGTAATGCTCCATC
	Tyrosinase-like	TRINITY_DN234089_c0_g1		AGAACGGTGGACGAGGTGT	GAGCGTGATGTTGGTGAAAG
	Tyrosinase-like	TRINITY_DN247553_c0_g1		GGTCCATTTTCAATTCGTGTG	GCTCCAGTAACATCAAGAGCAG
	Tyrosinase-like	TRINITY_DN294234_c0_g1		ATTGTTGGTGGGGAGATGTT	AGTCAAGGGTGATTTGGTCGT
	Tyrosinase-like	TRINITY_DN509330_c0_g4		CGCAGAGGAACAACTACATCAG	GAATACCTGGACTCATCGAACC
全缘冬青亚油酸代谢相关基因 linoleic acid metabolism related genes in I. integra	Linoleate 10R-lipoxygenase-like	TRINITY_DN22314_c0_g1		AACTCGGCTTACGGTCTTTTC	AACATTCCCTTGTCGTTCGT
	Linoleate 10R-lipoxygenase-like	TRINITY_DN75152_c0_g1		ACTGGGCGTTTAGTTGCTGT	GAGAGTCCGTCATCCGTGTC
	Linoleate 10R-lipoxygenase-like	TRINITY_DN81998_c0_g1		GACATTTACGCCTTCGTTTTCT	ATTTCTTCCTGGTCTGGCTTCT
	Psi-producing oxygenase A-like	TRINITY_DN173841_c0_g2		CAACATCCTTAGTTGCATGGTC	TCTTGAATCGTGACATCCGTAG
	Lipophospholipase-like	TRINITY_DN383657_c0_g1		TTTTTCTGGTGGTGGTATGAGA	TGAGCAGTTGTAGGCTTTACTCC
类别 type	基因 gene		基因号 gene ID	正向引物 forward primer (5'—3')	反向引物 reverse primer (5'—3')
全缘冬青花生四烯酸代谢相关基因arachidonic acid metabolism related genes in I. integra	Glutathione S-transferase		TRINITY_DN13210_c1_g1	AAGAGGTACTTGACCGAAAGGA	CGAAGATCACAGATATGGAAACC
	Glutathione S-transferase		TRINITY_DN23731_c0_g1	ATCTCAAGGCAAACAATGAAGG	CGATCTTTTCGTCTTTTCCAAC
	Glutathione S-transferase		TRINITY_DN25882_c0_g1	GCTGGATGGAACTTTTGGAAC	CACAGCTTTCTTCTCCTCATCTT
	Glutathione S-transferase		TRINITY_DN107276_c0_g2	GCTCCTGCTAAAAATGGCTTC	CCTCTTACTGAACAAAGGTTGATTG
	Glutathione S-transferase		TRINITY_DN306749_c0_g2	GTTATGTGCTTCCACCTGTTTG	GAGGAGATCCATTTGCTGAGAG
全缘冬青和红果冬青(I. p)过氧化氢酶基因 catalase genes in I. integra and I. purpurea (I. p)			TRINITY_DN40435_c0_g3	CGTGAATCTCCTGAACACAAAG	GAACACGATACCAATTTCTCACAG
	Catalase		TRINITY_DN47768_c1_g2	TCGTCAACTGTCGGTGTTTG	ACAGCATGGCTCTTGTTAATCC
			TRINITY_DN359401_c0_g1	AAGGTTTGGAATGACCAGCA	TGTTAGGACGGAAGCTCACAG
	Catalase(I. p)		TRINITY_DN25268_c0_g1	AACTGAATCTGGGCACAACC	TTCGAGAGGGGTAATTTGGA
			TRINITY_DN205218_c0_g1	AAGAGCTACCTGGAGGATGACC	ATGTGATGCCCTTGACACC
全缘冬青和红果冬青(I. p)内参基因 Internal control genes in I. integra and I. purpurea (I. p)	Actin			CCAGAGTCCAACACAATACCAG	GAGCACCCAGTTCTTCTCACA
	Actin(I. p)			CATCCCTTAGCACCTTCCAAC	ATCCTCACGTCAATGCTTCTC

表1

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NaCl处理下全缘冬青和红果冬青根系的转录组活性比较

Comparison of transcriptomic activity of Ilex integra and I. purpurea roots with NaCl treatments

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