外源IAA对马尾松幼苗茎干次生生长的影响

doi:10.11707/j.1001-7488.20210905

摘要/Abstract

摘要：

目的: 对2年生马尾松幼苗进行不同浓度IAA喷施处理，结合生理生化、形态解剖结构及转录组水平变化等综合分析，揭示外源IAA对马尾松苗期茎干次生生长的影响机制，为阐明马尾松苗期茎干次生生长的分子调控机制奠定基础，可以为培育速生高产的马尾松新品系提供一定的理论参考。方法: 对2年生马尾松幼苗进行不同浓度IAA（0、1、50、100 mg·L^-1）喷施处理，处理160天后进行生长指标、解剖结构、生理生化指标的测定；通过高通量转录组测序鉴定差异表达基因，对差异表达基因进行GO和KEGG功能分析。结果: IAA处理对2年生马尾松的地径、细胞生长（木质部、韧皮部厚度以及木质部细胞层数）、木材主要成分（木质素、纤维素及半纤维素含量）及内源激素含量（生长素、赤霉素及油菜素内酯）具有显著的促进作用（P＜0.05）；通过Illumina测序技术对对照组和IAA处理的6个cDNA文库进行测序，最终得到IAA处理后差异表达基因778个（包括482个上调表达基因和296个下调表达基因），GO分析表明这些差异基因主要富集于细胞成分、分子功能和生物过程中共计22个类别，KEGG分析表明它们参与了64种不同的途径，并筛选出与马尾松次生生长相关的差异表达基因（BRL1、GASA6、CAD和CESA2等）。结论: IAA处理能够改变内源激素含量，促进细胞分裂，增加木质素、纤维素和半纤维素积累，从而促进马尾松茎干的次生生长。

关键词: 马尾松, IAA, 次生生长, 转录组

Abstract:

Objective: Pinus massoniana is a unique fast-growing tree species in China and is one of the main afforestation tree species in southern China, which has high economic and ecological value. Auxin has a significant effect on the secondary growth of plants. An in-depth study on the secondary growth of the stem of P. massoniana can provide theoretical basis for breeding new varieties of P. massoniana with fast growth and high yield. In this study, two-year-old P. massoniana seedlings were sprayed with different concentrations of IAA. The effects of exogenous IAA on the secondary growth of P. massoniana seedlings were analyzed in combination with physiological, biochemical, morphological traits and anatomical structures and transcriptome changes. This study could lay a foundation for elucidating molecular regulation mechanism of stem secondary growth of P. massoniana seedlings. Method: Two-year-old P. massoniana seedlings were sprayed with different concentrations of IAA(0, 1, 50, 100 mg·L^-1). After 160 days of treatment, growth traits, anatomic structure, physiological and biochemical properties were measured. The differentially expressed genes were identified by high-throughput transcriptome sequencing, and analyzed by GO and KEGG. Result: IAA treatment significantly promoted the ground diameter, cell growth(xylem and phloem thickness and xylem cell layers), physiological components of wood(lignin, cellulose and hemicellulose), endogenous hormone contents(auxin, gibberellin and brassinolide) of two-year-old P. massoniana(P < 0.05). Six cDNA libraries of CK and IAA were sequenced by Illumina sequencing, 778 differentially expressed genes(including 482 up-regulated genes and 296 down-regulated genes) were obtained. GO analysis showed that these differential genes were mainly involved in 22 categories of cell components, molecular functions and biological processes, and KEGG analysis showed that they were involved in 64 different pathways. Differentially expressed genes related to secondary growth of P. massoniana were screened, such as BRL1, GASA6, CAD, CESA2, etc. Conclusion: IAA treatment changes the content of endogenous hormones, promotes cell division, increases the accumulation of lignin, cellulose and hemicellulose. Thus it promotes the secondary growth of P. massoniana. Our results provide a solid basis for revealing the mechanism of hormone regulation of secondary growth of P. massoniana.

Key words: Pinus massoniana, IAA, secondary growth, transcriptome

中图分类号:

周紫晶,范付华,尚先文,覃慧娟,王聪慧,丁贵杰,谭健晖. 外源IAA对马尾松幼苗茎干次生生长的影响[J]. 林业科学, 2021, 57(9): 42-51.

Zijing Zhou,Fuhua Fan,Xianwen Shang,Huijuan Qin,Conghui Wang,Guijie Ding,Jianhui Tan. Effects of Exogenous IAA on Stem Secondary Growth of Pinus massoniana Seedlings[J]. Scientia Silvae Sinicae, 2021, 57(9): 42-51.

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基因 Gene	序列 Sequence(5′→3′)
TRINITY_DN10179_c0_g1	F: TGATGGATGGTGGTTGATAC
	R: CTGATGGGCAAACTGAGATA
TRINITY_DN8564_c0_g2	F: GCGTATTCCGCCAGAGTG
	R: TCCAGAACGAGTGCCCCT
TRINITY_DN5190_c1_g1	F: CAGGCGACTCCACCTACAACT
	R: TGCGGAACACCTAATCCAAAC
TRINITY_DN2065_c0_g1	F: GAACGCAGAGCAATGAAG
	R: AAGCCCACCACTATGACC
TRINITY_DN11133_c0_g2	F: TCACGCTTCCTTTGGCTTTT
	R: CGTACGGTCTGCACTTCATC
TRINITY_DN3030_c1_g1	F: AGAGCAGATTCCAGCAGA
	R: CGGGTTTCCAGTAGATGA
TRINITY_DN39456_c0_g1	F: TCAGCGGACCCTGTTCTA
	R: GACAAAGTTGCCGTGGAG
TRINITY_DN40433_c0_g1	F: AGGGACTGGCTCCTTGTG
	R: CTTCGGAATCGCACCTTT
TRINITY_DN99422_c0_g3	F: GAAGACTTGAAATGCCTGAC
	R: ATAGACCTTGAACGCCTC
UBC	F: GATTTATTTCATTGGCAGGC
	R: AGGATCATCAGGATTTGGGT

IAA	木质素含量 Lignin content/(mg·g^-1)	纤维素含量 Cellulose content/(mg·g^-1)	半纤维素含量 Hemicellulose content/(mg·g^-1)
CK	89.95±0.98 a	199.30±35.81 a	160.51±1.01 a
1 mg·L^-1	116.33±0.07 b	292.35±18.03 b	207.13±15.40 b
50 mg·L^-1	122.43±3.92 b	287.18±16.06 b	202.68±5.75 b
100 mg·L^-1	124.94±7.08 b	285.85±12.47 b	187.36±3.05 ab

	内源激素Endogenous hormone/(ng·g^-1)	CK	IAA处理 IAA treatment
生长素类 Auxins	吲哚-3-乙酸Indole-3-acetic acid(IAA)	58.67±1.95	113.67±4.91^**
	氧化吲哚乙酸Oxidized indoleacetic acid(OxIAA)	0.00	114.00±4.73^**
	吲哚-3-甲醛Indole-3-carboxaldehyde(ICAld)	4.48±0.04	133.00±3.51^**
	吲哚-3-乙酸甲酯Indole-3-methyl acetate(MEIAA)	3.85±0.10	7.18±0.28^**
	吲哚乙酸-亮氨酸Indoleacetic acid-leucine(IAA-Leu)	1.06±0.08^**	0.56±0.03
	吲哚乙酸-天冬氨酸Indoleacetic acid-aspartic acid(IAA-Asp)	1.89±0.13^**	1.27±0.06
	色胺Tryptamine(TRA)	0.16±0.01	0.14±0.00
	色氨酸Tryptophan(TRP)	2 156.67±53.64^*	1 903.33±43.33
	3-吲哚丙酸3-Indolepropionic acid(IPA)	42.63±0.92	44.30±2.86
赤霉素类 Gibberellins	赤霉素1 Gibberellin1(GA1)	17.67±0.69	24.60±2.46
赤霉素类 Gibberellins	赤霉素15 Gibberellin15(GA15)	0.27±0.05	0.27±0.03
油菜素内酯类 Brassinolides	油菜素内酯Brassinolide(BR)	0.05±0.00	0.09±0.00^**
	油菜素甾酮Castasterone(CS)	19.89±1.18	27.03±0.99^**
	28-高油菜素甾酮28-homocastasterone(28-homoCS)	1.25±0.07	1.58±0.03^**
	香蒲甾醇Typhasterol(TY)	36.38±0.31	35.42±0.32
	6-脱氧油菜素甾酮6-deoxocastasterone(6-deoxoCS)	0.02±0.00	0.03±0.00

样本 Sample	原始读数 Raw reads	清洁读数(清洁读数/原始读数) Clean reads(clean reads/raw reads)	错误率 Error rate(%)	Q20(%)	Q30(%)	GC含量 GC content(%)
CK1	79 111 552	78 288 654(98.96%)	0.024 4	98.31	94.73	44.47
CK2	73 641 850	72 846 758(98.92%)	0.024 3	98.33	94.83	44.96
CK3	86 263 202	85 424 670(99.03%)	0.024 4	98.28	94.66	44.65
IAA1	87 975 948	87 084 030(98.99%)	0.024 2	98.36	94.87	44.83
IAA2	93 638 284	92 535 608(98.82%)	0.024 6	98.24	94.55	45.16
IAA3	77 429 912	76 588 978(98.91%)	0.024 7	98.17	94.39	44.87

基因号 Gene ID	注释 Description	差异倍数 Log₂FC	表达量Expression quantity
基因号 Gene ID	注释 Description	差异倍数 Log₂FC	CK	IAA
激素Hormone
TRINITY_DN6209_c0_g1	油菜素内酯受体激酶Brassinosteroid LRR receptor kinase(BRL1)	1.92	9.15	30.11
TRINITY_DN105600_c0_g2	赤霉素响应蛋白Gibberellin-regulated protein(GASA6)	1.70	2.09	6.35
木质素Lignin
TRINITY_DN40433_c0_g1	肉桂醇脱氢酶Cinnamyl alcohol dehydrogenases(CAD)	7.48	0	1.06
纤维素Cellulose
TRINITY_DN2065_c0_g1	纤维素合酶Cellulose synthase(CESA2)	1.13	23.30	45.01