水稻减数分裂期高温对苯丙烷类代谢及下游分支代谢途径的影响

doi:10.16819/j.1001-7216.2023.221112

中国水稻科学 ›› 2023, Vol. 37 ›› Issue (4): 368-378.DOI: 10.16819/j.1001-7216.2023.221112

水稻减数分裂期高温对苯丙烷类代谢及下游分支代谢途径的影响

汪胜勇¹, 陈宇航¹, 陈会丽¹, 黄钰杰¹, 张啸天¹, 丁双成²(), 王宏伟¹()

¹农业农村部长江中游作物绿色高效生产重点实验室(部省共建)/长江大学农学院，湖北荆州434025
²湿地生态与农业利用教育部工程研究中心/长江大学农学院，湖北荆州434025

收稿日期:2022-11-24 修回日期:2023-02-07 出版日期:2023-07-10 发布日期:2023-07-17
通讯作者: *email: shchding@yangtzeu.edu.cn;wanghw@yangtzeu.edu.cn
基金资助:
湿地生态与农业利用教育部工程研究中心开放基金资助项目(KFT202012);主要粮食作物产业化湖北省协同创新中心开放基金资助项目(KFT202108)

Effects of High Temperature on Phenylpropane Metabolism and Downstream Branch Metabolic Pathways in Rice Meiosis

WANG Shengyong¹, CHEN Yuhang¹, CHEN Huili¹, HUANG Yujie¹, ZHANG Xiaotian¹, DING Shuangcheng²(), WANG Hongwei¹()

¹MARA Key Laboratory of Sustainable Crop Production in the Middle Reaches of the Yangtze River (Co-construction by Ministry and Province)/College of Agriculture, Yangtze University, Jingzhou 434025, China
²Engineering Research Center of Ecology and Agricultural Use of Wetland, Ministry of Education/ College of Agriculture, Yangtze University, Jingzhou 434025, China

Received:2022-11-24 Revised:2023-02-07 Online:2023-07-10 Published:2023-07-17
Contact: *email: shchding@yangtzeu.edu.cn;wanghw@yangtzeu.edu.cn

摘要/Abstract

摘要：

【目的】 探究水稻减数分裂期高温如何影响苯丙烷类代谢，并分析其与水稻耐热性的关系。【方法】 以N22、广陆矮15、SDWG005、全两优681、Y两优900、Y两优1号、两优培九和绵恢101(MH101)等8种耐热和不耐热水稻品种为试验材料，设置常温和高温处理，分析减数分裂期高温胁迫对水稻的花粉活力与苯丙烷类代谢关键酶活性、木质素、总黄酮及总酚等主要代谢产物含量之间的相关性；并进一步选择极端耐高温的SDWG005和极端不耐高温的MH101为材料分析苯丙烷类代谢、碳水化合物代谢和抗氧化系统对水稻耐热性的影响。【结果】 1) 与对照相比，高温显著降低水稻花粉活力和颖花受精率，不同的水稻品种受高温影响后，花粉活力和颖花受精率的降幅不同。2) 高温显著增加颖花中肉桂酸-4-羟化酶和4-香豆酸辅酶A连接酶活性以及木质素、总黄酮和总酚的含量，且耐热品种增幅高于敏感品种。3) 高温下花粉活力与肉桂酸-4-羟化酶活性、木质素含量显著相关，颖花受精率与木质素含量以及木质素含量与类黄酮含量极显著相关。4) 与MH101相比，SDWG005小穗颖壳中木质素受高温显著诱导积累，且高温下能够维持较高细胞壁过氧化物酶活性。5) 与MH101相比，SDWG005颖花在高温下能够维持较高过氧化物酶、超氧化物歧化酶和抗坏血酸氧化酶活性，进而减少颖花中过氧化氢和丙二醛的积累。高温下SDWG005颖花中淀粉含量更高，酸性转化酶、蔗糖合酶及ATPase基因的表达量显著增加。【结论】 减数分裂期高温促进颖花中苯丙烷类代谢关键酶活性的上升和代谢产物含量的增加，耐热品种高温下能够积累较多的木质素和类黄酮，具有较高抗氧化酶活性，同时蔗糖代谢和能量产生效率较高，从而具有较强的耐热性。

关键词: 水稻, 高温, 颖花, 苯丙烷类代谢, 木质素, 总黄酮

Abstract:

【Objective】 Our purposes are to investigate how high temperature during rice meiosis affects phenylpropane metabolism and analyze its relationship with heat tolerance of rice. 【Methods】 Eight rice varieties differed in heat tolerance including N22, GLA15 (Guanglu’ai 15), SDWG005, QLY681 (Quanliangyou 681), YLY900 (Y Liangyou 900), YLY1 (Y Liangyou 1), LYP9 (Liangyoupeijiu), and MH101 (Mianhui 101) were used as experimental materials and exposed to room temperature and high temperature for 5 days. The correlations between activities of key enzymes in phenylpropane metabolism and main metabolite contents such as lignin, total flavonoids and total phenols were analyzed as well as the fertility of rice under high temperature stress during the meiotic stage. SDWG005 (heat resistant) and MH101 (heat sensitive) were used as experimental materials to analyze the effects of phenylpropane metabolism, carbohydrate metabolism and antioxidant defense system on heat tolerance of rice. 【Results】 1) Compared with the control, the pollen vitality and fertilization rate of spikelets decreased significantly to various extents at high temperature. 2) HT(high temperature) significantly increased the activities of cinnamate-4-hydroxylase and 4-coumaric acid coenzyme A ligase, as well as the accumulation of lignin, flavonoids and total phenols in spikelets, with resistant varieties registering a higher growth than sensitive ones. 3) Correlation analysis showed that in response to HT pollen activity was significantly correlated with cinnamate-4-hydroxylase activity and lignin content, spikelet fertility was significantly correlated with lignin content, and lignin content was significantly correlated with flavonoid content. 4) Compared with MH101, lignin accumulation in the glumes of SDWG005 was significantly induced under HT. And higher cell wall peroxidase activities were maintained in SDWG005 under HT. 5) Compared with MH101, SDWG005 could maintain higher antioxidant enzyme activities under HT, resulting in less accumulation of H₂O₂ and malondialdehyde. Under HT, the starch level in SDWG005 flowers was higher, and the expression levels of genes involved in acid invertase, sucrose synthase and ATPase were significantly upregulated. 【Conclusion】 High temperature stress increases the key enzyme activities and metabolite contents in the phenylpropanoid pathway in spikelets during the meiosis stage. The resistant variety accumulates more lignin and flavonoids during HT, has higher antioxidant enzyme activities, higher sucrose metabolism and energy utilization efficiency, thereby improving heat tolerance.

Key words: rice, high temperature, spikelet, phenylpropane metabolism, lignin, flavonoids

汪胜勇, 陈宇航, 陈会丽, 黄钰杰, 张啸天, 丁双成, 王宏伟. 水稻减数分裂期高温对苯丙烷类代谢及下游分支代谢途径的影响[J]. 中国水稻科学, 2023, 37(4): 368-378.

WANG Shengyong, CHEN Yuhang, CHEN Huili, HUANG Yujie, ZHANG Xiaotian, DING Shuangcheng, WANG Hongwei. Effects of High Temperature on Phenylpropane Metabolism and Downstream Branch Metabolic Pathways in Rice Meiosis[J]. Chinese Journal OF Rice Science, 2023, 37(4): 368-378.

图/表 8

图1 常温与高温人工气候室的温度设置

Fig. 1. Temperature in greenhouses for HT and CK.

表1 基因表达分析使用的引物

Table 1. Primers used in gene expression analysis.

基因位点号(基因名) Locus ID(gene name)	正向引物序列 Forward(5'-3')	反向引物 Reverse(5'-3')
LOC_Os09g08072(INV1)	AAGAGCAGGGGTGTACAAG	AAGCTGTGAGTCTGTGGCTC
LOC_Os06g09450(SUS2)	GTGTGCTTGACACCATCCAC	CATGCGGAGACAGGATAACA
LOC_Os05g45420(SnRK1A)	ACAACCAGTGGCTACCTTGG	CGATGATCAGTGGCTGAGTT
LOC_Os07g09610(SnRK1B)	ATATCAGGCGCCGAATACTG	TGTGCCTGAAGAACTTGCTG
LOC_Os05g14550(TOR)	GCTGAACGCTGCAATGACTA	ACCGAACAAGTACTGGAGCA
NC_001320.1(ATPase)	TCGGTGGAGCTACTCTTGGA	CGGGCGCGGATCTATGAATA
Tubulin	TACCGTGCCCTTACTGTTCC	CGGTGGAATGTCACAGACAC

图2 不同水稻品种减数分裂期高温处理下的花粉活力(A)和颖花受精率(B) GLA15－广陆矮15；QLY681－全两优681；LYP9－两优培九；YLY900－Y两优900；YLY1－Y两优1号；MH101－绵恢101。平均值±标准差，n = 3，柱状图上不同小写字母表示不同品种及处理间差异达5%显著水平。下同。

Fig. 2. Pollen vitality (A) and spikelet fertility (B) of different rice cultivars under HT during meiosis stage. GLA15, Guanglu’ai 15; QLY681, Quanliangyou 681; LYP9, Liangyoupeijiu; YLY900, Y Liangyou 900; YLY1, Y Liangyou 1; MH101, Mianhui 101. Mean ± standard deviation, n = 3; different lowercase letters on the bars indicate significant difference among varieties and treatments at 5% level. The same below.

图3 不同水稻品种减数分裂期高温处理下苯丙烷类代谢途径关键酶活性比较

Fig. 3. Comparison of key enzyme activities involved in phenylpropanoid pathway in different rice cultivars under HT during meiosis stage.

图4 不同水稻品种减数分裂期高温处理下颖花木质素、总黄酮和总酚含量的比较

Fig. 4. Comparison of lignin, flavonoids and total phenol contents in different rice cultivars under HT during meiosis stage.

表2 CK和HT处理下苯丙烷类代谢及下游分支代谢途径相关指标相关性分析

Table 2. Correlation analysis of various indexes involved in phenylpropane metabolism and downstream branch metabolic pathways under CK and HT.

相关系数 Correlation coefficient		花粉活力 Pollen vitality	颖花受精率 Spikelet fertility	苯丙氨酸解氨酶活性 Phenylalanine ammonia lyase activity	肉桂酸-4- 羟化酶活性 Cinnamic acid- 4-hydroxylase activity	4-香豆酸辅酶 A连接酶活性 4-Coumaric acid coenzyme A ligase activity	总酚 Total phenols	类黄酮 Flavonoids	木质素 Lignin
CK	花粉活力Pollen viability	1.000
	颖花受精率Spikelet fertility	0.615	1.000
	苯丙氨酸解氨酶活性 Phenylalanine ammonialyase activity	0.193	0.662	1.000
	肉桂酸-4-羟化酶活性 Cinnamic acid-4-hydroxylase activity	0.499	0.505	0.545	1.000
	4-香豆酸辅酶A连接酶活性 4-Coumaric acid coenzyme A ligase activity	0.406	0.634	0.910**	0.648	1.000
	总酚Total phenols	0.446	0.596	0.836**	0.840**	0.930**	1.000
	总黄酮Flavonoids	0.253	0.510	0.872**	0.412	0.832**	0.709*	1.000
	木质素Lignin	0.334	0.576	0.918**	0.632	0.904**	0.864**	0.960**	1.000
HT	花粉活力Pollen vitality	1.000
	颖花受精率Spikelet fertility	0.905**	1.000
	苯丙氨酸解氨酶活性 Phenylalanine ammonialyase activity	0.188	0.499	1.000
	肉桂酸-4-羟化酶活性 Cinnamic acid-4-hydroxylase activity	0.838**	0.694	0.068	1.000
	4-香豆酸辅酶A连接酶活性 4-Coumaric acid coenzyme A ligase activity	0.416	0.403	0.485	0.546	1.000
	总酚Total phenols	0.561	0.629	0.539	0.539	0.693	1.000
	总黄酮Flavonoids	0.562	0.588	0.659	0.406	0.733*	0.662	1.000
	木质素Lignin	0.775*	0.887**	0.649	0.598	0.590	0.668	0.860**	1.000

图5 水稻减数分裂期高温对颖花细胞壁过氧化物酶活性的影响柱状图上不同小写字母表示不同品种及处理间差异达5%显著水平。

Fig. 5. Effects of HT on cell wall peoxidase activities in spikelets during meiosis stage. Different lowercase letters above the bars indicate significant difference among different varieties and treatments at 5% level.

图6 减数分裂期高温对水稻颖花中抗氧化酶活性、过氧化氢、丙二醛和碳水化合物含量以及蔗糖代谢和能量水平相关基因表达的影响柱状图上不同小写字母表示不同品种及处理间差异达5%显著水平。

Fig. 6. Effects of HT on antioxidant enzyme activities, H2O2, malondialdehyde, carbohydrate contents, and gene expression involved in sucrose metabolic and energy level in spikelets during meiosis stage. Different lowercase letters above the bars indicate significant difference among different varieties and treatments at 5% level.

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水稻减数分裂期高温对苯丙烷类代谢及下游分支代谢途径的影响

Effects of High Temperature on Phenylpropane Metabolism and Downstream Branch Metabolic Pathways in Rice Meiosis

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