丛枝菌根真菌通过调节枳根系多胺提高抗旱性

doi:10.16420/j.issn.0513-353x.2022-1036

摘要/Abstract

摘要：

为明确丛枝菌根真菌是否以及如何调节多胺以提高柑橘的抗旱性，以枳（Poncirus trifoliata）实生苗为材料，接种丛枝菌根真菌——根内根孢囊霉（Rhizophagus intraradices），分析在正常水分和干旱胁迫下枳生长、叶片水势以及根系活性氧、多胺、多胺合成前体物质水平和多胺合成/分解酶活性的变化。干旱处理9周抑制了R. intraradices对根系的侵染，侵染率下降了16.85%。干旱处理也抑制了枳的生长和叶片水势，但R. intraradices显著缓解了这种抑制影响。接种R. intraradices增加了干旱胁迫下枳根系中胍基丁胺、S-腺苷-L-蛋氨酸、腐胺和尸胺含量，但也显著抑制精胺、亚精胺、L-精氨酸和L-鸟氨酸含量以及过氧化氢和超氧阴离子自由基水平。同时，接种R. intraradices还增加了各类多胺合成酶如精氨酸脱羧酶、鸟氨酸脱羧酶、精胺合成酶、亚精胺合成酶以及S-腺苷蛋氨酸脱羧酶活性，也同时提高了多胺分解酶如二胺氧化酶和多胺氧化酶的活性。此外，根系腐胺和尸胺均与超氧阴离子自由基显著负相关，而亚精胺与根系过氧化氢和超氧阴离子自由基显著正相关。因此，推断丛枝菌根真菌改变干旱胁迫下枳根系多胺水平以减轻活性氧爆发。

关键词: 柑橘, 抗旱性, 菌根, 腐胺, 活性氧

Abstract:

In order to clarify whether and how arbuscular mycorrhizal fungi regulate polyamines to enhance drought tolerance of citrus，trifoliate orange（Poncirus trifoliata）seedlings were inoculated with an arbuscular mycorrhizal fungus Rhizophagus intraradices，and the changes in growth，leaf water potential，reactive oxygen species levels，polyamines and polyamine precursors levels，and polyamine synthesis/cleavage enzyme activities were analyzed under well-watered and drought stress. The drought with nine weeks inhibited the colonization rate of R. intraradices in roots by 16.85%. Drought also significantly inhibited the growth and leaf water potential of trifoliate orange，while inoculation with R. intraradices considerably mitigated the inhibitive effect. Inoculation with R. intraradices significantly increased agmatine，S-adenosyl-L-methionine，putrescine，and cadaverine concentrations in roots under drought stress，along with the significant reduction in spermidine，spermine，L-arginine，L-ornithine，hydrogen peroxide，and superoxide anion radicals levels in roots. R. intraradices inoculation also distinctly elevated activities of various polyamine synthetases，such as arginine decarboxylase，ornithine decarboxylase，spermidine synthetase，spermine synthetase，and S-adenosyl methionine decarboxylase，and also increased activities of polyamine metabolism enzymes，such as diamine oxidase and polyamine oxidase. In addition，putrescine and cadaverine concentrations were significantly and negatively correlated with root superoxide anion radicals levels，while spermidine concentrations were positively correlated with root hydrogen peroxide and superoxide anion radicals levels. Therefore，it is concluded that arbuscular mycorrhizal fungi alter polyamine levels in roots of drought-stressed trifoliate orange to reduce reactive oxygen species burst.

Key words: citrus, drought tolerance, mycorrhiza, putrescine, reactive oxygen species

梁圣敏, 张菲, 吴强盛. 丛枝菌根真菌通过调节枳根系多胺提高抗旱性[J]. 园艺学报, 2023, 50(12): 2680-2688.

LIANG Shengmin, ZHANG Fei, WU Qiangsheng. Arbuscular Mycorrhizal Fungi Improve Drought Tolerance of Trifoliate Orange Seedlings by Regulating Root Polyamines[J]. Acta Horticulturae Sinica, 2023, 50(12): 2680-2688.

图/表 7

参考文献 30

[1]	Amiri R, Nikbakht A, Etemadi N. 2015. Alleviation of drought stress on rose geranium [Pelargonium graveolens (L.) herit.] in terms of antioxidant activity and secondary metabolites by mycorrhizal inoculation. Scientia Horticulturae, 197:373-380. doi: 10.1016/j.scienta.2015.09.062 URL
[2]	Bai X F, Dai L Q, Sun H M, Chen M T, Sun Y L. 2019. Effects of moderate soil salinity on osmotic adjustment and energy strategy in soybean under drought stress. Plant Physiology and Biochemistry, 139:307-313. doi: S0981-9428(19)30119-6 pmid: 30927693
[3]	Bhantana P, Rana M S, Sun X C, Moussa M G. 2021. Arbuscular mycorrhizal fungi and its major role in plant growth, zinc nutrition,phosphorous regulation and phytoremediation. Symbiosis, 84:19-37. doi: 10.1007/s13199-021-00756-6
[4]	Cheng S, Zou Y N, Kuča K, Hashem A, Abd_Allah E F, Wu Q S. 2021. Elucidating the mechanisms underlying enhanced drought tolerance in plants mediated by arbuscular mycorrhizal fungi. Frontiers in Microbiology, 12:809473. doi: 10.3389/fmicb.2021.809473 URL
[5]	Cheng Y, Ma W, Li X, Miao W, Zheng L, Cheng B. 2012. Polyamines stimulate hyphal branching and infection in the early stage of Glomus etunicatum colonization. World Journal of Microbiology and Biotechnology, 28:1615-1621. doi: 10.1007/s11274-011-0967-0 URL
[6]	Cvikrová M, Gemperlová L, Martincová O, Vanková R. 2013. Effect of drought and combined drought and heat stress on polyamine metabolism in proline-over-producing tobacco plants. Plant Physiology and Biochemistry, 73:7-15. doi: 10.1016/j.plaphy.2013.08.005 pmid: 24029075
[7]	Ding Y E, Zou Y N, Wu Q S, Kuča K. 2022. Mycorrhizal fungi regulate daily rhythm of circadian clock in trifoliate orange under drought stress. Tree Physiology, 42:616-628. doi: 10.1093/treephys/tpab132 URL
[8]	Ducros V, Ruffieux D, Belvabesnet H, Defraipont F, Berger F, Favier A. 2009. Determination of dansylated polyamines in red blood cells by liquid chromatography-tandem mass spectrometry. Analytical Biochemistry, 390:46-51. doi: 10.1016/j.ab.2009.04.007 pmid: 19364488
[9]	García-Tejero I, Romero-Vicente R, Jiménez-Bocanegra J A, Martínez-García G, Durán-Zuazo V H, Muriel-Ferná ndez. 2010. Response of citrus trees to deficit irrigation during different phenological periods in relation to yield,fruit quality,and water productivity. Agricultural Water Management, 97:689-699. doi: 10.1016/j.agwat.2009.12.012 URL
[10]	Hu Y B, Xie W, Chen B D. 2020. Arbuscular mycorrhiza improved drought tolerance of maize seedlings by altering photosystem II efficiency and the levels of key metabolites. Chemical and Biological Technologies in Agriculture, 7:20. doi: 10.1186/s40538-020-00186-4
[11]	Kapoor D, Singh S, Kumar V, Romero R, Prasad R, Singh J. 2019. Antioxidant enzymes regulation in plants in reference to reactive oxygen species (ROS) and reactive nitrogen species (RNS). Plant Gene, 19:100182.
[12]	Liang S M, Zheng F L, Abd_Allah E F, Muthuramalingam P, Wu Q S, Hashem A. 2021. Spatial changes of arbuscular mycorrhizal fungi in peach and their correlation with soil properties. Saudi Journal of Biological Sciences, 28:6495-6499. doi: 10.1016/j.sjbs.2021.07.024 URL
[13]	Liang S M, Zheng F L, Wu Q S. 2022. Elucidating the dialogue between arbuscular mycorrhizal fungi and polyamines in plants. World Journal of Microbiology and Biotechnology, 38:159. doi: 10.1007/s11274-022-03336-y
[14]	Liu Jian-xin, Hu Hao-bin, Zhao Guo-lin. 2008. The relationship between the release of ethylene and the concentration of polyamines and the accumulation of reactive oxygen in Peganum multisectum Bobr leaves under drought stress. Acta Ecologica Sinica, 28 (4):1579-1585. (in Chinese)
	刘建新, 胡浩滨, 赵国林. 2008. 干旱胁迫下多裂骆驼蓬(Peganum multisectum Bobr)叶片乙烯释放和多胺含量变化与活性氧积累的关系. 生态学报, 28 (4):1579-1585.
[15]	Nandy S, Das T, Tudu C K, Mishra T. 2022. Unravelling the multi-faceted regulatory role of polyamines in plant biotechnology,transgenics and secondary metabolomics. Applied Microbiology and Biotechnology, 106:905-929. doi: 10.1007/s00253-021-11748-3
[16]	Nehela Y, Killiny N. 2020. The unknown soldier in citrus plants: polyamines-based defensive mechanisms against biotic and abiotic stresses and their relationship with other stress-associated metabolites. Plant Signaling & Behavior, 15 (6):1761080.
[17]	Ortas I, Rafique M,Çekiç F Ö. 2021. Do mycorrhizal fungi enable plants to cope with abiotic stresses by overcoming the detrimental effects of salinity and improving drought tolerance? //Symbiotic Soil Microorganisms. Cham:Springer:391-428.
[18]	Phillips M, Hayman D S. 1970. Improved procedures for clearing roots and staining parasitic and vesicular-arbuscular mycorrhizal fungi for rapid assessment of infection. Transactions of the British Mycological Society, 55:158-161. doi: 10.1016/S0007-1536(70)80110-3 URL
[19]	Shi J, Fu X Z, Peng T, Huang X S, Fan Q J, Liu J H. 2010. Spermine pretreatment confers dehydration tolerance of citrus in vitro plants via modulation of antioxidative capacity and stomatal response. Tree Physiology, 30:914-922. doi: 10.1093/treephys/tpq030 URL
[20]	Tan M, Hassan M J, Peng Y, Feng G Y, Huang L K, Liu L, Liu W, Han L B, Li Z. 2022. Polyamines metabolism interacts with γ-aminobutyric acid,proline and nitrogen metabolisms to affect drought tolerance of creeping bentgrass. International Journal of Molecular Sciences, 23 (5):2779. doi: 10.3390/ijms23052779 URL
[21]	Vashisth T, Chun C P, Hampton M O. 2020. Florida citrus nursery trends and strategies to enhance production of field-transplant ready citrus plants. Horticulturae, 6:8. doi: 10.3390/horticulturae6010008 URL
[22]	Velikova V, Yordanov I, Edreva A. 2000. Oxidative stress and some antioxidant systems in acid rain-treated bean plants:protective role of exogenous polyamines. Plant Science, 151:59-66. doi: 10.1016/S0168-9452(99)00197-1 URL
[23]	Wang Ai-guo, Luo Guang-hua. 1990. Quantitative relation between the reaction of hydroxylamine and superoxide anion radicals in plants. Plant Physiology Communications, 6:55-57. (in Chinese)
	王爱国, 罗广华. 1990. 植物的超氧物自由基与羟胺反应的定量关系. 植物生理学通讯, 6:55-57.
[24]	Wang Xue-kui. 2006. Principle and technology of plant physiological and biochemical experiment. Beijing: High Education Press. (in Chineses)
	王学奎. 2006. 植物生理生化实验原理和技术. 北京: 高等教育出版社.
[25]	Wu Q S, He J D, Srivastava A K, Zou Y N, Kuca K. 2019. Mycorrhizas enhance drought tolerance of citrus by altering root fatty acid compositions and their saturation levels. Tree Physiology, 39:1149-158. doi: 10.1093/treephys/tpz039 URL
[26]	Wu Q S, Srivastava A K, Zou Y N. 2013. AMF-induced tolerance to drought stress in citrus:a review. Scientia Horticulturae, 164:77-87. doi: 10.1016/j.scienta.2013.09.010 URL
[27]	Zhang F, Zou Y N, Wu Q S. 2018. Quantitative estimation of water uptake by mycorrhizal extraradical hyphae in citrus under drought stress. Scientia Horticulturae, 229:132-136. doi: 10.1016/j.scienta.2017.10.038 URL
[28]	Zhang F, Zou Y N, Wu Q S, Kuča K. 2020. Arbuscular mycorrhizas modulate root polyamine metabolism to enhance drought tolerance of trifoliate orange. Environmental and Experimental Botany, 171:103926. doi: 10.1016/j.envexpbot.2019.103926 URL
[29]	Zhao J, Wang X, Pan X, Jiang Q, Xi Z. 2021. Exogenous putrescine alleviates drought stress by altering reactive oxygen species scavenging and biosynthesis of polyamines in the seedlings of Cabernet Sauvignon. Frontiers in Plant Science, 14:767992.
[30]	Zou Y N, Wu Q S, Kuča K. 2021. Unravelling the role of arbuscular mycorrhizal fungi in mitigating the oxidative burst of plants under drought stress. Plant Biology, 23:50-57. doi: 10.1111/plb.v23.s1 URL

处理 Treatment	干旱胁迫前 Pre-drougth stress			干旱胁迫后 Post-drought stress
处理 Treatment	侵染率/% Colonization	株高/cm Height	茎粗/mm Stem diameter	侵染率/% Colonization	株高/cm Height	茎粗/mm Stem diameter
-AMF + WW	0.00 ± 0.00 b	5.55 ± 0.91 c	1.91 ± 0.22 b	0.00 ± 0.00 c	13.58 ± 0.69 c	2.14 ± 0.22 b
+AMF + WW	54.07 ± 7.85 a	17.83 ± 3.13 a	2.78 ± 0.24 a	84.97 ± 7.24 a	27.91 ± 3.18 a	3.38 ± 0.24 a
-AMF + DS	0.00 ± 0.00 b	4.07 ± 1.11 c	1.89 ± 0.12 b	0.00 ± 0.00 c	11.11 ± 0.83 d	2.04 ± 0.12 b
+AMF + DS	55.05 ± 8.27 a	15.01 ± 1.37 b	2.73 ± 0.23 a	70.65 ± 3.45 b	24.99 ± 1.44 b	3.31 ± 0.23 a

处理 Treatment	干旱胁迫前 Pre-drougth stress			干旱胁迫后 Post-drought stress
处理 Treatment	侵染率/% Colonization	株高/cm Height	茎粗/mm Stem diameter	侵染率/% Colonization	株高/cm Height	茎粗/mm Stem diameter
-AMF + WW	0.00 ± 0.00 b	5.55 ± 0.91 c	1.91 ± 0.22 b	0.00 ± 0.00 c	13.58 ± 0.69 c	2.14 ± 0.22 b
+AMF + WW	54.07 ± 7.85 a	17.83 ± 3.13 a	2.78 ± 0.24 a	84.97 ± 7.24 a	27.91 ± 3.18 a	3.38 ± 0.24 a
-AMF + DS	0.00 ± 0.00 b	4.07 ± 1.11 c	1.89 ± 0.12 b	0.00 ± 0.00 c	11.11 ± 0.83 d	2.04 ± 0.12 b
+AMF + DS	55.05 ± 8.27 a	15.01 ± 1.37 b	2.73 ± 0.23 a	70.65 ± 3.45 b	24.99 ± 1.44 b	3.31 ± 0.23 a

处理 Treatment	生物量/（g · plant^-1）Biomass		根冠比 Root/shoot ratio	叶片水势/MPa Leaf water potential
处理 Treatment	地上部 Shoot	地下部 Root	根冠比 Root/shoot ratio	叶片水势/MPa Leaf water potential
-AMF + WW	0.86 ± 0.06 c	1.31 ± 0.12 c	1.54 ± 0.17 a	-0.43 ± 0.02 bc
+AMF + WW	2.07 ± 0.18 a	2.22 ± 0.29 a	1.07 ± 0.14 b	-0.36 ± 0.02 a
-AMF + DS	0.75 ± 0.07 c	1.04 ± 0.10 d	1.40 ± 0.17 a	-0.45 ± 0.03 c
+AMF + DS	1.83 ± 0.31 b	1.81 ± 0.24 b	0.99 ± 0.10 b	-0.40 ± 0.03 b

处理 Treatment	生物量/（g · plant^-1）Biomass		根冠比 Root/shoot ratio	叶片水势/MPa Leaf water potential
处理 Treatment	地上部 Shoot	地下部 Root	根冠比 Root/shoot ratio	叶片水势/MPa Leaf water potential
-AMF + WW	0.86 ± 0.06 c	1.31 ± 0.12 c	1.54 ± 0.17 a	-0.43 ± 0.02 bc
+AMF + WW	2.07 ± 0.18 a	2.22 ± 0.29 a	1.07 ± 0.14 b	-0.36 ± 0.02 a
-AMF + DS	0.75 ± 0.07 c	1.04 ± 0.10 d	1.40 ± 0.17 a	-0.45 ± 0.03 c
+AMF + DS	1.83 ± 0.31 b	1.81 ± 0.24 b	0.99 ± 0.10 b	-0.40 ± 0.03 b

处理 Treatment	L-精氨酸 L-Arg	L-鸟氨酸 L-Orn	胍基丁胺 Agm	S-腺苷-L-蛋氨酸 SAM
-AMF + WW	353.4 ± 37.0 d	7.73 ± 0.72 d	0.32 ± 0.02 d	1.01 ± 0.07 c
+AMF + WW	564.6 ± 31.8 a	20.60 ± 2.07 a	2.45 ± 0.11 a	3.18 ± 0.17 a
-AMF + DS	504.5 ± 17.8 b	16.71 ± 1.15 b	0.61 ± 0.05 c	1.16 ± 0.06 c
+AMF + DS	401.2 ± 18.5 c	10.13 ± 1.27 c	1.24 ± 0.10 b	1.79 ± 0.08 b