印度梨形孢对水稻的促生作用及其机理的初探

doi:10.3969/j.issn.1001-7216.2015.02.012

摘要/Abstract

摘要：

印度梨形孢(Piriformospora indica)是一种能与植物建立互惠共生关系的根部内生真菌,能促进植物生长,增加作物产量。为了研究印度梨形孢对水稻生长发育的影响,将印度梨形孢与籼稻苗共培养后,分析印度梨形孢对水稻植株高度、生物量、分蘖数、抽穗期、穗粒性状、叶绿素含量、硝酸还原酶活性、根系活力以及生长相关调控基因表达水平的影响。结果显示,接种印度梨形孢的水稻株高、地上部干鲜质量、分蘖数、硝酸还原酶活性和根系活力显著高于对照;与对照相比,印度梨形孢定殖的水稻植株抽穗期提前4~6 d;印度梨形孢处理第3、4周后,叶片叶绿素含量比对照分别增加了19.77%和17.89%。RT-PCR分析表明,印度梨形孢定殖的水稻植株叶片中生长素相关调控基因OsIAA13和YUCCA的上调表达,表达量分别为对照的1.49倍和1.30倍;负调控水稻生长的NRR基因表达量为对照的58%。印度梨形孢促进水稻地上部的生长与叶绿素含量、硝酸还原酶活性、根系活力和生长相关调控基因的表达有关,印度梨形孢可能通过提高光合速率、增强水稻对矿质营养的吸收与利用和诱导生长素的分泌,促进水稻地上部的生长。

关键词: 印度梨形孢, 水稻, 叶绿素, 硝酸还原酶, 根系活力, 生长素

Abstract:

Piriformospora indica is a root endophytic fungus, which is able to establish beneficial symbiotic relationships with plant, and it can promote the growth of plants, enhance crop yield. In order to study the impact of P. indica on growth and development of rice, We analyzed the differences in plant height, biomass, tillering, heading, panicle properties, chlorophyll content, nitrate reductase (NR) activity, root activity and expression of growth-related genes between nonglutinous rice colonized and un-colonized by P. indica. The results showed that the plant height, biomass of shoot, the number of tillers, root activity and NR activity of rice colonized by P. indica were significantly higher than that in un-colonized ones. Rice colonized by P. indica headed 4-6 days earlier than the un-colonized rice. The contents of chlorophyll increased by 19.77% and 17.89% respectively after P. indica treatment for 3 and 4 weeks. RT-PCR analysis showed that the expression of auxin-regulated genes OsIAA13 and YUCCA were up-regulated in leaves of rice colonized by P. indica, being 1.49 and 1.30 folds as high as that of control plants. The expression of NRR, which negatively regulates the growth of rice, was 58% as high as that of control plants. The results indicated that the growth promotion of rice conferred by P. indica was relevant to chlorophyll content, NR activity, root activity and expression of growth-related genes. P. indica may promote the growth of rice through improving photosynthetic rate, enhancing mineral nutrition absorption and utilization and inducing auxin secretion.

Key words: Piriformospora indica, rice, chlorophyll, nitrate reductase, root activity, auxin

中图分类号:

吴金丹, 陈乾, 刘晓曦, 林福呈, 高其康, 楼兵干. 印度梨形孢对水稻的促生作用及其机理的初探[J]. 中国水稻科学, 2015, 29(2): 200-207.

Jin-dan WU, Qian CHEN, Xiao-xi LIU, Fu-cheng LIN, Qi-kang GAO, Bing-gan LOU. Preliminary Study on Mechanisms of Growth Promotion in Rice Colonized by Piriformospora indica[J]. Chinese Journal OF Rice Science, 2015, 29(2): 200-207.

图/表 11

图2 印度梨形孢对水稻幼苗生长的影响 A-接种印度梨形孢20 d后(左为对照,右为根部接种印度梨形孢);B-接种印度梨形孢40 d后(左为对照,右为根部接有印度梨形孢)。

Fig. 2. Influence of P. indica on rice seedling growth. A, 20 days after inoculation with P. indica (Left: control; Right: colonized by P. indica); B, 40 days after inoculation with P. indica (Left: control; Right: colonized by P. indica).

表1 生长相关基因 qRT-PCR特异性引物

Table 1 Specific primers of growth-related genes for quantitative RT-PCR.

基因 Gene	引物序列 Primer sequences
OsIAA13	5'-CCGCAACTGGGAACAATATG-3'
	5'-ACAAGCATCCAGTCACCATC-3'
YUCCA	5'-GATGTGTTCGGTGAAATGCC-3'
	5'-AGCGCCTATGGTTCAGTAATC-3'
NRR	5'-AAGTGTCAGTTCTGGGTGC-3'
	5'-AACCTGAATGTCTCCTTCGC-3'
OsUBQ5^[19]	5'-GAAGGAGGAGGAAATCGAAC-3'
(内参基因)	5'-CTTCAC AGAGGTGATGCTAAGG-3'

图1 印度梨形孢在水稻根部的定殖箭头所示为印度梨形孢的厚垣孢子。

Fig. 1. Colonization of P. indica in rice root. Arrows indicate the chlamydospores of P. indica.

表2 印度梨形孢对水稻生物学性状的影响

Table 2 Effect of P. indica on biological characteristics of rice.

生物学性状 Biological characteristics	对照 Control			P. indica处理 P. indica treatment
株高 Plant height /cm	16.39	±	1.44 b B	17.79	±	0.89 a A
根长 Root length/cm	10.73	±	1.97 a A	12.05	±	2.37 a A
根数 Number of roots	10.33	±	1.54 a A	10.20	±	1.15 a A
最大叶长 Max leaf length/cm	9.96	±	1.13 b B	12.13	±	0.99 a A
最大叶宽 Max leaf width /cm	0.25	±	0.03 b B	0.30	±	0.02 a A
根鲜质量 Root weight/g	0.0300	±	0.0062 b A	0.0350	±	0.0063 a A
地上部鲜质量 Shoot weight/g	0.0720	±	0.0121 b B	0.0860	±	0.0154 a A
根干质量 Dry root weight/g	0.0036	±	0.0008 a A	0.0041	±	0.0006 a A
地上部干质量 Dry shoot weight/g	0.0091	±	0.0015 b B	0.0111	±	0.0016 a A

图3 印度梨形孢对水稻分蘖数的影响 -P.indica-对照; +P.indica-接种印度梨形孢。

Fig. 3. Effect of P. indica on number of tillers in rice. -P.indica, Control; +P.indica, Inoculation with P. indica.

图4 印度梨形孢对水稻抽穗期的影响 -P.indica-对照; +P.indica-接种印度梨形孢。

Fig. 4. Effect of P. indica on rice heading. -P.indica, Control; +P.indica, Inoculation with P. indica.

表3 印度梨形孢对水稻穗粒性状的影响

Table 3 Effect of P. indica on rice panicle properties.

穗粒性状 Panicle property	岳优9113 Yueyou 9113		钱优2号 Qianyou 2
穗粒性状 Panicle property	-P. indica	+P. indica	-P. indica	+P. indica
穗长 Length of panicle/cm	20.33 b B	22.34 a A	18.39 b B	20.38 a A
每穗总粒数 Total grains per panicle	80.92 b B	111.16 a A	79.84 b A	96.52 a A
每穗实粒数 Filled grains per panicle	64.12 b B	86.84 aA	56.64 b B	69.04 a A
结实率 Seed-setting rate/%	79.24 a A	78.12 a A	70.94 a A	71.53 a A
千粒重 1000-grain weight/g	21.62 a A	22.51 a A	23.53 a A	23.81 a A

图5 印度梨形孢对水稻叶片叶绿素含量的影响 -P. indica -对照; +P. indica -接种印度梨形孢。

Fig. 5. Effect of P. indica on chlorophyll content in rice leaf. -P. indica, Control; +P. indica, Inoculation with P. indica.

图6 印度梨形孢对硝酸还原酶活性的影响 -P. indica -对照; +P. indica -接种印度梨形孢。

Fig. 6. Effect of P. indica on nitrate reductase activity. -P. indica, Control; +P. indica, Inoculation with P. indica.

图7 印度梨形孢对根系活力的影响 -P. indica -对照; +P. indica -接种印度梨形孢。

Fig. 7. Effect of P. indica on root activity. -P. indica, Control; +P. indica, Inoculation with P. indica.

图8 印度梨形孢对生长相关基因表达水平的影响 -P. indica -对照; +P. indica -接种印度梨形孢。

Fig. 8. Effect of P. indica on growth-related genes expression levels. -P. indica, Control; +P. indica, Inoculation with P. indica.

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