内源激素含量变化以及与激素合成相关基因表达量对毕克齐山药块茎膨大的影响

敖兰吉亚; 季祥; 邵盈; 赵令敏; 张艳芳; 霍秀文

内源激素含量变化以及与激素合成相关基因表达量对毕克齐山药块茎膨大的影响

1.
内蒙古农业大学园艺与植物保护学院/内蒙古野生蔬菜种质资源与创新重点实验室，内蒙古　呼和浩特　010019

基金项目: 国家自然科学基金项目（31860558）

详细信息

作者简介:
敖兰吉亚（1994−），女，硕士研究生，研究方向：蔬菜遗传育种与生物技术（E-mial：1140842979@qq.com）

通讯作者:
霍秀文（1968−），女，教授，博士生导师，研究方向为：蔬菜遗传育种与生物技术（E-mail：huoxiuwen@imau.edu.cn.com）

中图分类号: S 632.1
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出版历程
- 收稿日期: 2020-01-06
- 修回日期: 2020-03-05
- 网络出版日期: 2020-05-30

Effect of endogenous hormone content and gene expression related to hormone synthesis on tuber expansion of Bikeqi yam

1.
College of Horticulture and Plant Protection, Inner Mongolia Agricultural University/Key Laboratory of Wild Vegetable Germplasm Resources and Innovation, Hohhot, Inner Mongolia,　010019

摘要

摘要: 目的通过分析毕克齐山药中内源激素含量与淀粉含量、可溶性总糖含量和还原糖含量的相关性，以及与其相关合成基因表达量的相关性分析，探究毕克齐山药块茎膨大的机理。方法以毕克齐山药5个不同生长期块茎为材料，采用酶联免疫吸附法分别测定脱落酸、赤霉素、生长素、茉莉酸、玉米素、异戊稀基腺苷这6种内源激素含量，采用高效液相色谱仪测定方法测定水杨酸含量。结果内源激素生长素（IAA）、玉米素（ZR）、脱落酸（ABA）、茉莉酸（JA）和水杨酸（SA）含量与山药块茎形态指标正相关；内源激素赤霉素（GA3）、异戊稀基腺苷（IPA）含量与形态指标负相关；内源激素IAA含量与山药块茎周长和块茎直径显著正相关；内源激素GA3含量与块茎长度显著负相关；与IAA相关的基因与内源激素IAA含量显著负相关结论内源激素IAA、ZR、ABA、JA和SA促进山药块茎膨大；内源激素GA3、IPA抑制山药块茎生长；内源激素IAA促进山药增粗；内源激素GA3抑制其伸长生长；与IAA相关的基因的下调表达对IAA的合成有促进作用，即正调控内源激素IAA含量。
- 山药 /
- 块茎膨大 /
- 形态指标 /
- 生理指标 /
- 内源激素 /
- 基因表达量
Abstract: Objective To investigate the changes of starch content, reducing sugar content and soluble total sugar content in tubers, the changes of endogenous hormone content in tubers, the morphological indexes, physiological indexes and the analysis of variance of endogenous hormones during the process of tuber expansion. And the correlation between these indicators and endogenous hormones, lay the foundation for further study of the physiological mechanism of yam growth and development and guiding the high yield and quality cultivation of yam. Method Six kinds of endogenous hormones such as ABA, GA3, IAA, JA, ZR and IPA were determined by enzyme-linked immunosorbent assay with different tuber lengths of Bikeqi yam. High performance liquid chromatography (HPLC) was used to determine the content of salicylic acid (SA), and to analyze the dynamic changes of source hormone content in different growth stages, and the correlation between morphological indicators of yam and endogenous hormones. The relationship between differentially expressed genes associated with endogenous hormones and endogenous hormones was explored for the effects of these factors on the expansion of yam tubers. Result The contents of IAA, ZR, ABA, JA and SA were positively correlated with the morphological indexes of yam tuber; the contents of GA3 and IPA were negatively correlated with the morphological indexes; the contents of IAA were positively correlated with the perimeter and diameter of yam tuber; the contents of GA3 were negatively correlated with the length of tuber; the genes correlated with IAA were negatively correlated with the contents of IAA Conclusion Endogenous hormones IAA, ZR, ABA, JA and SA promote the expansion of yam tuber; endogenous hormones GA3 and IPA inhibit the growth of yam tuber; endogenous hormones IAA promote the thickening of yam; endogenous hormones GA3 inhibit the elongation of yam; the down-regulation of IAA related genes can promote the synthesis of IAA, that is, regulate the content of IAA.
- yam /
- tuber enlargement /
- morphological index /
- physiological index /
- endogenous hormone /
- endogenous hormone related gene

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图 1 生长发育过程中各个生理指标的变化

A-生长发育过程中还原糖含量的变化A-The change of reducing sugar content in the process of growth and development B-生长发育过程中可溶性总糖含量的变化B-Changes in reducing sugar content during growth and development C-生长发育过程中淀粉含量的变化C-Changes in starch content during growth and development

Figure 1. Changes of physiological indexes in the process of growth and development

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表 1 毕克齐山药在块茎膨大过程中内源激素含量变化

Table 1. Changes of endogenous hormones in BIkeqi yam varieties during tuber enlargement

	105 d	120 d	135 d	150 d	165 d
IAA	24.697±1.82 b	38.542±2.15 b	46.454±3.46 a	37.793±2.41 ab	48.012±2.68 a
ZR	5.295±0.23 b	6.97±0.47 b	8.129±0.45 ab	9.39±0.72 a	6.751±0.36 ab
ABA	33.274±1.93 bc	33.564±0.03 c	28.402±1.70 bc	40.707±2.11 b	81.939±1.88 a
JA	11.976±0.43 a	12.678±0.48 c	15.806±1.02 b	13.541±0.92 b	21.276±1.46 b
GA₃	5.484±0.44 a	2.538±0.11 ef	4.072±0.40 bc	3.209±0.20 cde	3.69±0.26 bc
IPA	4.103±0.20 a	8.005±0.37 a	5.943±0.37 a	4.202±0.21 a	4.419±0.27 a
SA	52.308±30.38 b	88.763±63.26 ab	19.226±10.83 b	81±37.48 ab	234.867±111.44 a
注：不同小写字母表示在p＜0.05水平上差异显著。 Note: Different lowercase letters indicate significant differences at the p＜0.05 level.

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表 2 毕克齐山药内源激素与形态、生理指标相关性

Table 2. Correlation between endogenous hormones and morphological and physiological indicators in Bikeqi Yam

	块茎长度 Length	块茎周长 Circumference	块茎直径 Diameter	块茎鲜重 Fresh weight	块茎干重 Dry weight	淀粉含量 Starch content	还原糖含量 Reducing sugar content	可溶性总糖含量 Total soluble sugar content	IAA	ZR	ABA	JA	GA3	IPA	SA
块茎长度 Length		0.756	0.757	0.674	0.679	0.375	−0.78	−0.940*	0.841	0.562	0.431	0.531	−0.892*	0.435	0.494
块茎周长 Circumference			1.000**	0.922*	0.981**	0.719	−0.823	−0.839	0.906*	0.589	0.64	0.833	−0.425	−0.161	0.553
块茎直径 Diameter				0.924*	0.982**	0.715	−0.822	−0.838	0.904*	0.588	0.643	0.833	−0.425	−0.162	0.557
块茎鲜重 Fresh weight					0.972**	0.489	−0.84	−0.75	0.692	0.657	0.665	0.694	−0.43	−0.362	0.591
块茎干重 Dry weight						0.646	−0.817	−0.771	0.807	0.602	0.677	0.804	−0.36	−0.321	0.579
淀粉含量 Starch content							−0.617	−0.611	0.767	0.542	0.072	0.524	−0.061	−0.04	−0.082
还原糖含量 Reducing sugar content								0.927*	−0.732	−0.932*	−0.21	−0.375	0.673	−0.034	−0.17
可溶性总糖含量 Total soluble sugar content									−0.883*	−0.773	−0.289	−0.494	0.805	−0.315	−0.293
IAA										0.483	0.479	0.779	−0.533	0.227	0.433
ZR											−0.116	0.043	−0.575	0.022	−0.159
ABA												0.871	−0.118	−0.378	0.975**
JA													−0.107	−0.242	0.787
GA3														−0.612	−0.252
IPA															−0.222
在0.05级别，相关性显著。在0.01级别，相关性极显著。：Significant correlation（P＜0.05）；**：Extremely significant correlation（P＜0.01）.

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表 3 与内源激素合成相关基因的基因编号、基因代号和基因注释信息

Table 3. Gene number, gene code and predictive gene of genes involved in endogenous hormone synthesis

与IAA合成相关基因 Genes related to IAA synthesis	基因ID Gene ID	基因代号 Gene code	基因注释信息 Gene annotation information
	Unigene0016848	IAA 1	生长素响应蛋白基因 Auxin response protein gene
	Unigene0021936	IAA 2	生长素响应蛋白基因 Auxin response protein gene
	Unigene0023040	IAA 3	生长素响应蛋白基因 Auxin response protein gene
	Unigene0025388	IAA 4	IAA-氨基酸水解酶ILR1基因 IAA-amino acid hydrolase ILR1 gene
	Unigene0027417	IAA 5	IAA-氨基酸水解酶ILR1基因 IAA-amino acid hydrolase ILR1 gene
	Unigene0027794	IAA 6	生长素响应蛋白基因 Auxin response protein gene
	Unigene0028701	IAA 7	生长素响应蛋白基因 Auxin response protein gene
	Unigene0028916	IAA 8	生长素响应蛋白基因 Auxin response protein gene
	Unigene0030426	IAA 9	生长素响应蛋白基因 Auxin response protein gene
与GA₃合成相关基因 Genes related to GA₃ synthesis	Unigene0002430	GA₃ 1	赤霉素2-β-双加氧酶 Gibberellin 2-β-dioxygenase
	Unigene0019924	GA₃ 2	赤霉素2-β-双加氧酶 Gibberellin 2-β-dioxygenase
	Unigene0021502	GA₃ 3	赤霉素2-β-双加氧酶 Gibberellin 2-β-dioxygenase
	Unigene0021503	GA₃ 4	赤霉素2-β-双加氧酶 Gibberellin 2-β-dioxygenase
	Unigene0027812	GA₃ 5	赤霉素20氧化酶 Gibberellin 2-β-dioxygenase
	Unigene0023907	GA₃ 6	赤霉素调节蛋白基因 Gibberellin regulatory protein gene
	Unigene0024440	GA₃ 7	赤霉素调节蛋白基因 Gibberellin regulatory protein gene
	Unigene0026445	GA₃ 8	赤霉素调节蛋白基因 Gibberellin regulatory protein gene
	Unigene0020765	GA₃ 9	赤霉素调节蛋白基因 Gibberellin regulatory protein gene

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表 4 与内源激素合成相关基因的rpkm平均值

Table 4. rpkm mean of genes associated with endogenous hormone synthesis

与IAA合成相关基因 Genes related to IAA synthesis	基因 IDGene ID	基因代号 Gene code	105 d	120 d	135 d	150 d	165 d
	Unigene0016848	IAA 1	95.82±30.63	65.29±44.90	64.25±22.21	26.73±12.83	26.68±10.96
	Unigene0021936	IAA 2	14.49±2.45	8.08±6.19	3.64±2.22	2.95±3.33	3.57±0.87
	Unigene0023040	IAA 3	213.97±40.34	119.15±61.21	132.16±54.901	48.88±36.07	39.56±9.26
	Unigene0025388	IAA 4	4.61±3.19	5.45±0.97	8.84±2.12	1.18±0.84	0
	Unigene0027417	IAA 5	11.82±1.17	7.21±3.44	5.10±1.42	1.48±1.15	3.18±0.68
	Unigene0027794	IAA 6	2.28±0.33	0.82±0.67	0.53±0.42	0.28±0.39	0
	Unigene0028701	IAA 7	60.78±12.10	30.43±21.84	77.84±20.86	50.67±17.82	21.64±5.46
	Unigene0028916	IAA 8	127.67±26.48	40.53±21.48	66.59±24.93	35.91±11.76	14.06±1.25
	Unigene0030426	IAA 9	7.79±1.10	2.82±3.12	3.91±1.72	0.36±0.51	2.20±0.80
与GA₃合成相关基因 Genes related to GA₃ synthesis	Unigene0002430	GA₃ 1	24.60±5.23	24.72±10.21	18.71±6.22	5.74±6.31	2.92±0.83
	Unigene0019924	GA₃ 2	4.80±1.80	0.62±0.60	1.20±1.54	0	0
	Unigene0021502	GA₃ 3	4.02±1.56	1.21±1.06	3.05±2.99	2.34±1.78	0
	Unigene0021503	GA₃ 4	5.82±3.34	5.51±2.87	5.20±2.06	6.88±7.74	0
	Unigene0027812	GA₃ 5	13.12±2.80	1.38±1.23	2.59±0.96	1.91±0.92	5.69±1.14
	Unigene0023907	GA₃ 6	16.76±10.56	4.20±5.94	0.67±0.60	0.08±0.12	0.42±0.60
	Unigene0024440	GA₃ 7	176.89±19.66	189.53±72.98	110.84±19.67	80.80±39.49	423.10±68.62
	Unigene0026445	GA₃ 8	47.20±12.01	32.18±34.09	51.94±10.28	74.13±97.99	203.55±60.52
	Unigene0020765	GA₃ 9	11.95±2.51	1.74±1.95	4.61±2.35	0.55±0.78	0

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表 5 内源激素IAA含量与其合成相关基因表达量的相关性分析

Table 5. Correlation analysis between endogenous hormone IAA and its synthesis-related gene expression

	IAA含量 IAA content	IAA 1	IAA 2	IAA 3	IAA 4	IAA 5	IAA 6	IAA 7	IAA 8	IAA 9
IAA含量 IAA content		−0.681	−0.875	−0.719	−0.046	−0.744	−0.902^*	−0.230	−0.799	−0.644
IAA 1			0.868	0.992^**	0.669	0.956^*	0.912^*	0.482	0.898^*	0.916^*
IAA 2				0.869	0.218	0.961^**	0.966^**	0.128	0.833	0.869
IAA 3					0.638	0.944^*	0.936^*	0.547	0.946^*	0.936^*
IAA 4						0.442	0.340	0.700	0.481	0.453
IAA 5							0.944^*	0.245	0.855	0.937^*
IAA 6								0.373	0.944^*	0.904^*
IAA 7									0.643	0.418
IAA 8										0.911^*
IAA 9
在0.05级别，相关性显著。在0.01级别，相关性极显著。：Significant correlation（P＜0.05）；**：Extremely significant correlation（P＜0.01）.

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表 6 内源激素GA₃含量与其合成相关基因表达量的相关性分析

Table 6. Correlation analysis between endogenous hormone GA₃ and its synthesis-related gene expression

	GA₃含量 GA₃ content	GA₃ 1	GA₃ 2	GA₃ 3	GA₃ 4	GA₃ 5	GA₃ 6	GA₃ 7	GA₃ 8	GA₃ 9
GA₃含量 GA₃ content		0.256	0.863	0.680	0.020	0.900^*	0.737	−0.007	−0.041	0.872
GA₃ 1			0.652	0.557	0.511	0.267	0.648	−0.400	−0.803	0.673
GA₃ 2				0.765	0.306	0.876	0.958^*	−0.185	−0.416	0.991^**
GA₃ 3					0.728	0.468	0.620	−0.730	−0.704	0.826
GA₃ 4						−0.096	0.277	−0.957^*	−0.910^*	0.354
GA₃ 5							0.873	0.224	0.058	0.827
GA₃ 6								−0.085	−0.383	0.914^*
GA₃ 7									0.854	−0.265
GA₃ 8										−0.469
GA₃ 9
在0.05级别，相关性显著。在0.01级别，相关性极显著。：Significant correlation（P＜0.05）；**：Extremely significant correlation（P＜0.01）.

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