丛枝菌根真菌通过上调根系及自身水孔蛋白基因表达提高玉米抗旱性

doi:10.3724/SP.J.1258.2012.00973

摘要/Abstract

摘要：

在模拟干旱条件下, 研究了接种丛枝菌根(AM)真菌Glomus intraradices对玉米(Zea mays)根部13种质膜水孔蛋白基因表达的影响, 同时观测了AM真菌自身水孔蛋白基因的表达情况。结果表明, 干旱条件下, 除Zm PIP1;3、Zm PIP1;4、Zm PIP1;5和Zm PIP2;2之外的接种处理能显著提高根部其他8种质膜水孔蛋白基因的表达(Zm PIP2;7表达量未检测出), 并且AM真菌菌丝中水孔蛋白基因GintAQP1表达也显著增强。与此同时, 接种处理明显改善了植物水分状况, 提高了叶片水势。AM真菌增强宿主植物根部及自身的水孔蛋白基因的表达对于提高植物抗旱性具有潜在的重要贡献。

关键词: 水孔蛋白, AM真菌, 抗旱性, 叶片水势, 玉米

Abstract:

Aims It has been well demonstrated that arbuscular mycorrhizal (AM) fungi can improve water balance and drought tolerance of host plants under drought stress. However, controversy still exists in mechanisms underlying the mycorrhizal functions. For example, in different experiments AM fungi could up- or down-regulate plant aquaporin gene expression. Furthermore, little information is available on the expression of aquaporin genes in AM fungi under drought stress and its contribution to plant drought tolerance. We investigated the effects of an AM fungus, Glomus intraradices, on expression of a plasma membrane intrinsic protein (PIP) gene family containing 13 PIP genes in maize roots and one aquaporin gene from the AM fungus under simulated drought conditions. Our objectives were to systematically investigate the aquaporin gene expression in the mycorrhizal association in response to drought stress and to help understand the molecular basis for drought tolerance of AM symbiosis.
Methods Maize plants inoculated with/without AM fungus G. intraradices were grown under different water regimes in a controlled-environment climate chamber for 42 days. At harvest, the leaf water potential (Ψ) was determined with an SKPM 1400 pressure chamber, and then shoots and roots were collected and carefully cleaned with tap water. Three grams of mixed roots were used to estimate the percentage root colonization after clearing with 10% KOH and staining with 0.05% (v/v) trypan blue in lactic acid. One gram of mixed roots was used to extract total RNA by using TRIZOL to synthesize cDNA. Real-time PCR analysis was performed to estimate the expression of Zm PIP genes and GintAQP1. Shoots and the rest of roots were dried at 105 °C for 10 min and 80 °C for 48 h to obtain dry weights. About 0.2 g of dried shoots or roots was digested by HNO₃for 12 h. The P concentration was recorded by using microplate reader spectra at the wavelength of 820 nm. Experimental data were subjected to statistical analysis using SPSS 13.0.
Important findings Under drought stress, inoculation with AM fungus significantly enhanced expression of eight PIP genes except for Zm PIP1;3, Zm PIP1;4 Zm PIP1;5 and Zm PIP2;2 (Zm PIP2;7 expression not detected), and drought stress also enhanced expression of GintAQP1 cloned from hyphae of G. intraradices. Enhanced aquaporin gene expression was beneficial to improvement of plant water status and increase of leaf water potential.

Key words: aquaporin, arbuscular mycorrhizal (AM) fungus, drought tolerance, leaf water potential, maize

李涛, 陈保冬. 丛枝菌根真菌通过上调根系及自身水孔蛋白基因表达提高玉米抗旱性. 植物生态学报, 2012, 36(9): 973-981. DOI: 10.3724/SP.J.1258.2012.00973

LI Tao, CHEN Bao-Dong. Arbuscular mycorrhizal fungi improving drought tolerance of maize plants by up-regulation of aquaporin gene expressions in roots and the fungi themselves. Chinese Journal of Plant Ecology, 2012, 36(9): 973-981. DOI: 10.3724/SP.J.1258.2012.00973

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链接本文: https://www.plant-ecology.com/CN/10.3724/SP.J.1258.2012.00973

https://www.plant-ecology.com/CN/Y2012/V36/I9/973

图/表 8

表1 不同水分条件下玉米根系菌根侵染率(平均值±标准偏差, n = 4)

Table 1 Mycorrhizal colonization rate of maize roots under different water regimes (mean ± SD, n = 4)

接种处理 Inoculation treatment	菌根侵染率 Mycorrhizal colonization rate (%)
接种处理 Inoculation treatment	正常水分 Well-watered	干旱 Drought
不接种 Non-inoculated	0.0	0.0
接种 Inoculated	35.1^*	38.5^{* #}

图1 不同水分条件下AM真菌Glomus intraradices对玉米植株干重的影响(平均值±标准偏差, n = 4)。分别对地上部和根部进行双因素方差分析和t检验。*表示相同水分处理条件下不同接种处理间在5%水平上差异显著。

Fig. 1 Effects of inoculation with arbuscular mycorrhizal (AM) fungus Glomus intraradices on maize dry weights under different water regimes (mean ± SD, n = 4). Two-way ANOVA and t-test were performed for shoots and roots, respectively. * indicates significant difference (p < 0.05) between inoculation treatments under the same water treatment.

图2 不同水分条件下AM真菌Glomus intraradices对玉米植株磷浓度的影响(平均值±标准偏差, n = 4)。分别对地上部和根部进行双因素方差分析和t检验。*表示相同水分处理条件下不同接种处理间在5%水平上差异显著。

Fig. 2 Effects of inoculation with arbuscular mycorrhizal (AM) fungus Glomus intraradices on P concentration of maize under different water regimes (mean ± SD, n = 4). Two-way ANOVA and t-test were performed for shoots and roots, respectively. * indicates significant difference (p < 0.05) between inoculation treatments under the same water treatment.

表2 地上部和根部干重、磷浓度及叶片水势的双因素(接种处理和水分处理)方差分析

Table 2 Two-way ANOVA of shoot and root dry weight, P concentration and leaf water potential as influenced by inoculation and water treatments

	地上部干重 Shoot dry weight	根部干重 Root dry weight	地上部磷含量 Shoot P concentration	根部磷含量 Root P concentration	叶片水势 Leaf water potential
接种处理 Inoculation treatment (I)	*	ns	*	ns	*
水分处理 Water treatment (W)	ns	ns	ns	ns	**
交互作用 I × W interaction	ns	ns	ns	ns	ns

图3 不同水分条件下AM真菌Glomus intraradices对玉米叶片水势的影响(平均值±标准偏差, n = 4)。#表示相同接种处理下不同水分处理间在5%水平上差异显著。

Fig. 3 Effects of inoculation with arbuscular mycorrhizal (AM) fungus Glomus intraradices on leaf water potential of maize under different water regimes (mean ± SD, n = 4). # indicates significant difference (p < 0.05) between water treatments under the same inoculation treatment.

图4 不同水分条件下AM真菌Glomus intraradices对玉米根部质膜水孔蛋白基因表达的影响(平均值±标准偏差, n = 4)。#表示在相同接种处理下不同水分处理间在5%水平差异显著。*表示在相同水分处理条件下不同接种处理间在5%水平上差异显著。对不同基因分别进行统计分析。D, 干旱处理; W, 正常水分。

Fig. 4 Effects of arbuscular mycorrhizal (AM) fungus Glomus intraradices on expression of plasma membrane aquaporin gene in maize roots under different water regimes (mean ± SD, n = 4). # indicates significant difference (p < 0.05) between water treatments under the same inoculation treatment. * indicates significant difference (p < 0.05) between inoculation treatments under the same water treatment. Statistical analysis was performed for each aquaporin gene respectively. D, drought treatment; W, well-watered treatment.

表3 12个质膜水孔蛋白基因表达的双因素(接种处理和水分处理)方差分析

Table 3 Two-way ANOVA of gene expression of 12 Zm PIPs as influenced by inoculation and water treatments.

	Zm PIP1;1	Zm PIP1;2	Zm PIP1;3	Zm PIP1;4	Zm PIP1;5	Zm PIP1;6	Zm PIP2;1	Zm PIP2;2	Zm PIP2;3	Zm PIP2;4	Zm PIP2;5	Zm PIP2;6
接种处理 Inoculation treatment (I)	*	**	ns	ns	ns	**	**	**	**	*	**	**
水分处理 Water treatment (W)	**	**	ns	ns	*	ns	*	ns	ns	ns	**	ns
交互作用 I ×W interaction	ns	ns	ns	ns	ns	ns	ns	ns	ns	ns	ns	ns

图5 干旱胁迫对AM真菌Glomus intraradices水孔蛋白基因表达的影响(平均值±标准偏差, n = 4)。#表示在相同接种处理下不同水分处理间在5%水平差异显著。

Fig. 5 Effects of drought stress on expression of arbuscular mycorrhizal (AM) fungus Glomus intraradices aquaporin gene (mean ± SD, n = 4). # indicates significant difference (p < 0.05) between water treatments under the same inoculation treatment.

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