丛枝菌根真菌对低磷胁迫下核桃幼苗根系磷吸收的影响及机制

doi:10.11707/j.1001-7488.LYKX20220253

摘要/Abstract

摘要：

目的: 探索低磷（P）胁迫下丛枝菌根（AM）真菌对核桃幼苗根系P吸收的影响及机制，为核桃幼苗的栽培生产提供参考。方法: 基于前期筛选的1个高效AM真菌菌株——沾屑多孢囊霉，分析适P（100 μmol·L^?1）和低P（1 μmol·L^?1）条件下沾屑多孢囊霉对‘辽河一号’核桃幼苗植株生长、根系构型、根系P含量、根系和土壤磷酸酶活性以及根系酸性磷酸酶分泌基因（JrPAP10和JrPAP12）和磷转运基因（JrPT3;1和JrPT3;2）相对表达量的影响。结果: 沾屑多孢囊霉接种3个月后，核桃幼苗根系侵染率达45.6%~53.2%，且低P处理可提高根系菌根侵染。在低P和适P条件下，接种沾屑多孢囊霉显著提高植物生物量和土壤酸性、中性和碱性磷酸酶活性；低P条件下沾屑多孢囊霉还显著改善植物根系构型，提高根系P含量以及根系酸性磷酸酶活性。根系JrPAP10和JrPT3;1均被沾屑多孢囊霉诱导表达，JrPAP12仅在低P条件时被沾屑多孢囊霉诱导表达，且JrPT3;2的表达量被沾屑多孢囊霉抑制。结论: AM真菌菌株沾屑多孢囊霉诱导低P条件下核桃幼苗JrPAP10、JrPAP12和JrPT3;1基因的表达，可促进土壤和根系磷酸酶活性，改善根系构型，从而提高核桃幼苗P吸收和生物量积累。

关键词: 核桃, 菌根, 磷, 磷酸酶, 磷转运基因

Abstract:

Objective: Walnut is a high-quality economic forest tree. Arbuscular mycorrhizal (AM) fungi can promote plant growth and phosphorus (P) uptake, however, the mechanisms by which AM fungi enhance nutrient utilization and plant growth in walnuts are not clear. This study aims to explore the effect of AM fungi on P uptake of walnuts under low P conditions and the associated mechanisms. Method: In this study, an effective AM fungal strain, Diversispora spurca, screened earlier, was used to inoculate Juglans regia cv. Liaohe 1 seedlings, and the effects of D. spurca on plant growth, root system architecture, root P, root and soil phosphatase activities, and the relative expression of root acid phosphatase secretion genes (JrPAP10 and JrPAP12) and phosphate transporter protein genes (JrPT3;1 and JrPT3;2) of J. regia cv. Liaohe 1 seedlings under appropriate P (100 μmol·L^?1) and low P (1 μmol·L^?1) level conditions were studied. Result: After three months of inoculation with D. spurca, the mycorrhizal colonization of walnut root system reached 45.6%–53.2%, and low P treatment significantly increased root mycorrhizal colonization, compared with appropriate P treatment. Inoculation of D. spurca significantly increased plant biomass and soil acid, neutral, and alkaline phosphatase activities under both low and appropriate P levels conditions. D. spurca also significantly improved root morphology as well as increased root P content and root acid phosphatase activity under low P condition. Root JrPAP10 expression was up-regulated by D. spurca under both low and appropriate P levels conditions, while JrPAP12 expression was induced by D. spurca only under low P level condition. In addition, D. spurca up-regulated the relative expression of JrPT3;1 in roots under both low P and appropriate P level conditions, while the expression of JrPT3;2 was inhibited by D. spurca. Conclusion: The AM fungus, D. spurca, can induce up-regulated expressions of JrPAP10, JrPAP12, and JrPT3;1 in roots of walnut seedlings under low P levels conditions, increase soil phosphatase, root acid phosphatase, and improve root morphology, and thereby enhancing P uptake and biomass accumulation of walnut seedlings.

Key words: walnut, mycorrhiza, phosphorus, phosphatase, phosphate transporter gene

中图分类号:

曹明奡,张菲,黄光明,刘瑞成,刘利平,吴强盛,徐永杰. 丛枝菌根真菌对低磷胁迫下核桃幼苗根系磷吸收的影响及机制[J]. 林业科学, 2023, 59(12): 117-124.

Ming'ao Cao,Fei Zhang,Guangming Huang,Ruicheng Liu,Liping Liu,Qiangsheng Wu,Yongjie Xu. Effects of Arbuscular Mycorrhizal Fungi on Phosphorus Uptake of Walnut Seedling Roots under Low Phosphorus Stress and the Potential Mechanisms[J]. Scientia Silvae Sinicae, 2023, 59(12): 117-124.

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基因 Genes	登记号 Accessions	引物序列 Primer sequence（5’→3’）
JrPAP10	Jm3DG00060700	F: ACACCCCTCAATACCAATGGC
		R: ACTCTCATGGTTTCCCCCTCC
JrPAP12	Jm4DG00150700	F: ACACTCCACAATACAAATGGC
		R: TGGGCAATGCATAAAAACGAT
JrPHT3;1	Jm2DG00002500	F: ATCTCATCTGGGTTTGGAGTG
		R: CTGAGCACTGTAGCCAAGCAA
JrPHT3;2	Jm6DG00001600	F: TGCCCACTTTTCTAGGTTACA
		R: AGGTTTTGTACTTGGTTGCAT
18S rRNA	LOC109010173	F: GGTCAATCTTCTCGTTCCCTT
		R: TCGCATTTCGCTACGTTCTT

AM真菌接种 AM fungal inoculation	P处理 P treatment	地上部生物量 Shoot biomass/（g·plant^?1）	地下部生物量 Root biomass/（g·plant^?1）	总生物量 Total biomass/（g·plant^?1）
?A	?P	7.74±0.47d	9.05±0.69d	18.11±1.39d
?A	+P	9.86±0.52c	12.03±1.25c	24.07±2.51c
+A	?P	12.11±1.08b	13.72±0.42b	27.43±0.84b
+A	+P	15.43±1.02a	15.32±1.13a	30.63±2.27a
显著性 Significance
A		0.0001	<0.0001	<0.0001
P		0.0001	0.0004	0.0005
A×P		0.1688	0.1665	0.1455

AM真菌接种 AM fungal inoculation	P处理 P treatment	总长 Total length/cm	投影面积 Projected area/cm²	表面积 Surface area/cm²	平均直径 Average diameter/mm	体积 Volume/cm³
?A	?P	600±65b	47.6±4.3b	149±16b	0.73±0.04c	3.05±0.16b
	+P	609±70b	58.2±6.7a	167±15ab	0.97±0.06a	3.09±0.25b
+A	?P	728±65a	60.3±4.0a	190±13a	0.94±0.09ab	4.42±0.25a
	+P	645±38ab	53.1±7.3ab	183±21a	0.85±0.09b	4.57±0.39a
显著性 Significance
A		0.0199	0.2068	0.0047	0.2377	<0.0001
P		0.2489	0.5673	0.5240	0.0775	0.4998
A×P		0.1560	0.0091	0.1622	0.0007	0.7064

变量 Variables	A	P	A×P	变量 Variables	A	P	A×P
土壤酸性磷酸酶Soil acid phosphatase	<0.0001	0.7781	0.0061	JrPT3;1	<0.0001	0.0002	0.0002
土壤中性磷酸酶Soil acid phosphatase	<0.0001	<0.0001	<0.0001	JrPT3;2	0.0002	<0.0001	0.0062
土壤碱性磷酸酶Soil alkaline phosphatase	<0.0001	<0.0001	<0.0001	JrPAP10	<0.0001	<0.0001	<0.0001
根系酸性磷酸酶Root acid phosphatase	0.0189	0.0350	0.2499	JrPAP12	0.7294	0.0256	0.0095
根系P Root P	<0.0001	<0.0001	<0.0001

项目	土壤磷酸酶 Soil acid phosphatase			根系酸性磷酸酶 Root acid phosphatase	根系P Root P	JrPT3;1	JrPT3;2	JrPAP10	JrPAP12	根系性状 Root character
项目	酸性 Acid	中性 Neutral	碱性 Alkaline	根系酸性磷酸酶 Root acid phosphatase	根系P Root P	JrPT3;1	JrPT3;2	JrPAP10	JrPAP12	根长 Root length	投影面积 Projected area	表面积 Surface area	平均直径 Diameter	体积 Volume
根系P Root P	0.57	0.40	0.67*	0.11	1.00	0.27	?0.93**	0.13	0.66*	0.22	0.58*	0.59*	0.61*	0.49
菌根侵染率 Mycorrhizal colonization	0.84**	0.90**	0.77**	0.51	0.45	0.85**	?0.48	0.71**	0.10	0.60*	0.33	0.67**	0.23	0.91**