灌浆充实调节剂对大穗型水稻弱势籽粒灌浆的调控效应

doi:10.7668/hbnxb.20192499

摘要/Abstract

摘要：

为探究2种灌浆充实调节剂对大穗型水稻品种弱势籽粒灌浆充实的影响及其作用机制,采用大田试验,以大穗型水稻品种交源优216为试验材料,研究了抽穗开花期喷施灌浆充实调节剂(禾立丰和新美洲星)对水稻弱势籽粒灌浆充实、内源激素含量、灌浆充实相关miRNA及其靶基因、编码灌浆充实相关蛋白和蔗糖-淀粉代谢关键酶基因表达量的影响。结果表明,2种灌浆充实调节剂禾立丰和新美洲星显著提高了交源优216的产量、弱势籽粒千粒质量、灌浆速率及灌浆起始势,弱势籽粒千粒质量与对照(清水处理)相比分别提高了16.07%和15.89%;且弱势籽粒中IAA含量在花后9,15,21 d显著增加,Z+ZR含量在花后9,21 d显著增加。禾立丰处理下,弱势籽粒中负调控籽粒灌浆充实的miR167a-c、miR167d-j和miR1432在花后6 d显著下调,编码14-3-3蛋白的基因OsGF14b和OsGF14f在花后12 d显著下调;而正调控籽粒灌浆充实的类萌发素蛋白基因OsGLP3在花后6 d显著上调,蔗糖-淀粉代谢关键酶基因在花后6,12 d显著上调。新美洲星处理下,弱势籽粒中负调控籽粒灌浆充实的miR167a-c、miR167d-j、miR1432在花后6 d显著下调,编码14-3-3蛋白基因OsGF14b和OsGF14f在花后12 d显著下调;正调控籽粒灌浆充实的OsGLP3和蔗糖-淀粉代谢关键酶基因的表达量在花后6,12 d显著上调。相关分析结果表明,弱势籽粒灌浆速率与miR1432、OsGF14f的表达量呈显著负相关关系,而与IAA含量呈显著正相关关系。因此,禾立丰和新美洲星可能是通过调节灌浆前中期灌浆充实相关miRNA及靶基因、编码灌浆充实相关蛋白和蔗糖-淀粉代谢关键酶基因的表达量,进而提高激素IAA和Z+ZR含量,促进弱势籽粒灌浆充实。

关键词: 水稻, 弱势籽粒, 灌浆充实, 调节剂, miRNA, 蔗糖-淀粉代谢关键酶基因

Abstract:

In order to explore the effects and mechanism of grain filling regulators on inferior spikelets filling of large panicle rice varieties,large panicle rice variety Jiaoyuanyou 216 was selected as material and an open field experiment was conducted by spraying exogenous grain filling regulators(Helifeng and Xinmeizhouxing)at heading stage.Meanwhile,endogenous hormone contents,expressions of filling-related miRNAs and their targets,and genes encoding filling-related proteins and sucrose-starch metabolism key enzymes were measured.The results should that the two grain filling regulators of Helifeng and Xinmeizhouxing significantly increased the yield of Jiaoyuanyou 216,the 1000-grain weight,filling rate and initial grain filling potential of inferior spikelets.Specifically,compared with the control(spraying water),the 1000-grain weight of inferior spikelets increased by 16.07% and 15.89%,respectively.Content of IAA in inferior spikelets increased significantly at 9,15,21 d after flowering,and the content of Z+ZR increased significantly at 9,21 d after flowering.Under Helifeng treatment,expressions of miR167a-c,miR167d-j and miR1432 were significantly down-regulated at 6 d after flowering,OsGF14b and OsGF14f were significantly down-regulated at 12 after flowering. What's more,expression of OsGLP3,a gene encoding germin-like protein was significantly up-regulated at 6 d after flowering,and expressions of sucrose-starch metabolism key enzyme related genes were significantly up-regulated at 6,12 d after flowering.However,under Xinmeizhouxing treatment,expressions of miR167a-c,miR167d-j,miR1432 were significantly down-regulated at 6 d after flowering,and expressions of OsGF14b and OsGF14f were significantly down-regulated at 12 d after flowering.More importantly,expressions of OsGLP3 and sucrose-starch metabolism key enzyme related genes were significantly up-regulated at 6,12 d after flowering.Correlation analysis results showed that the filling rate of inferior spikelets was significantly negatively correlated with the expression of miR1432 and OsGF14f,while was significantly positively correlated with the content of IAA.Therefore,Helifeng and Xinmeizhouxing may regulate the expressions of filling-related miRNAs and their targets,as well as the genes of filling-related proteins and sucrose-starch metabolism key enzymes in the pre-and mid-filling period and increase the content of IAA and Z+ZR to promote inferior spikelets filling.

Key words: Rice, Inferior spikelets, Grain filling, Regulators, miRNA, Sucrose-starch metabolism key enzyme related genes

静莉丽, 彭廷, 赵亚帆, 赵帅兵, 王童童, 李源, 程远, 杜彦修, 张静, 孙红正, 赵全志. 灌浆充实调节剂对大穗型水稻弱势籽粒灌浆的调控效应[J]. 华北农学报, 2022, 37(1): 86-94. doi: 10.7668/hbnxb.20192499.

JING Lili, PENG Ting, ZHAO Yafan, ZHAO Shuaibing, WANG Tongtong, LI Yuan, CHENG Yuan, DU Yanxiu, ZHANG Jing, SUN Hongzheng, ZHAO Quanzhi. Regulatory Effects of Two Grain Filling Regulators on Inferior Spikelets Filling of Large Panicle Rice[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(1): 86-94. doi: 10.7668/hbnxb.20192499.

http://www.hbnxb.net/CN/Y2022/V37/I1/86

图/表 9

表1 引物序列

Tab.1 Primer sequence

引物名称 Primer name	引物序列(5'—3') Primer sequence
miR167a-c_RT	CTCAACTGGTGTCGTGGAGTCGGCAATTCAGTTGAGTAGATCAT
miR167d-j_RT	CTCAACTGGTGTCGTGGAGTCGGCAATTCAGTTGAGCAGATCAT
miR1432_RT	CTCAACTGGTGTCGTGGAGTCGGCAATTCAGTTGAGGTCGGTGT
miR167a-c_F	ACACTCCAGCTGGGTGAAGCTGCCAGCAT
miR167d-j_F	ACACTCCAGCTGGGTGAAGCTGCCAGCAT
miR1432_F	ACACTCCAGCTGGGATCAGGAGAGATGAC
Stem-loop_U	TGGTGTCGTGGAGTCG
OsARF12_F	ACGATGTCACAATGCATGCG
OsARF12_R	AGTAATTCGTCGGCTGCTTG
OsACOT_F	AGGTTGAGGGAAAGCTGTTG
OsACOT_R	GGGCCTGCGCTATCAATTTG
OsGLP3_F	TGTCCAAGGTGCTCAACAAG
OsGLP3_R	TATCCACCTGAAACGCCTTC
OsGF14f_F	AGCAGCTGAGAACACTCTTG
OsGF14f_R	CAGCAATAGCATCGTCGAAC
OsGF14b_F	GCTTGAATCCCACCTTGTC
OsGF14b_R	AATGTCCTGAGCAGCCTTG
OsSUS3_F	CCCCAACTCCGACCTTTACT
OsSUS3_R	TTGCAATCAGGTCAGCTGTG
OsAGPS2_F	GTTCGTTGAAGTCCTCGCTG
OsAGPS2_R	TGCGGTAAAGGTGATCTCCA
OsGBSSI_F	AAGGTTGCAGACAGGTACGA
OsGBSSI_R	TCTTCTCACCGGTCTTTCCC
β-actin_F	GGAAGTACAGTGTCTGGATTGGTG
β-actin_R	TCTTGGCTTAGCATTCTTGGGT

表2 灌浆充实调节剂对水稻产量构成因素和产量的影响

Tab.2 Effects of the grain filling regulators on rice yield components and yield

处理 Treatment	穗数/ (×10⁴/hm²) No.of spikelets	穗粒数 Spikelets per panicle	结实率/% Seed setting percentage	千粒质量/g 1000-grain weight		产量/ (t/hm²) Yield
处理 Treatment	穗数/ (×10⁴/hm²) No.of spikelets	穗粒数 Spikelets per panicle	结实率/% Seed setting percentage	强势籽粒 Superior spikelets	弱势籽粒 Inferior spikelets	产量/ (t/hm²) Yield
CK	299.27±4.97a	284.07±9.39a	82.94±1.01c	27.46±0.30ab	16.93±0.38b	11.14±0.14b
T1	299.31±3.83a	289.48±9.46a	89.90±0.73a	27.87±0.16a	19.65±0.31a	11.70±0.17a
T2	299.25±4.66a	292.79±8.44a	87.00±0.98b	27.13±0.20b	19.62±0.86a	11.65±0.13a

图1 灌浆充实调节剂对水稻强、弱势籽粒千粒质量及灌浆速率的影响

Fig.1 The effects of the grain filling regulators on the 1000-grain weight and the grain filling rate of superior and inferior spikelets of rice

表3 灌浆充实调节剂处理下水稻弱势籽粒灌浆过程的Richards方程特征参数

Tab.3 Grain-filling Richards equation parameters for inferior spikelets of rice under the grain filling regulators

处理 Treatment	灌浆起始势 R₀	最大灌浆速率/(mg/d) V_max	平均灌浆速率/(mg/d) V	活跃灌浆期/d AGP
CK	0.073±0.000b	0.568±0.001c	0.364±0.001c	46.361±0.179a
T1	0.078±0.001a	0.708±0.002a	0.449±0.001a	41.332±0.174c
T2	0.077±0.000a	0.658±0.000b	0.419±0.000b	44.029±0.062b

图2 灌浆充实调节剂对水稻弱势籽粒中miRNA(A、B、D)及其靶基因表达量(C、E)的影响
不同字母代表同一开花后时间不同处理间的差异显著(P<0.05)。图3—5同。

Fig.2 Effects of the grain filling regulators on the level of miRNAs(A,B,D)and their targets expression(C,E)in inferior spikelets of rice
The different letters represent the significant difference among different treatments in the same days after flowering(P<0.05).The same as Fig.3—5.

图3 灌浆充实调节剂对水稻弱势籽粒灌浆启动关键信号蛋白基因表达量的影响

Fig.3 Effects of the grain filling regulators on the gene expression level of key signal proteins of filling initiation in inferior spikelets of rice

图4 灌浆充实调节剂对水稻弱势籽粒中蔗糖-淀粉代谢关键酶基因表达量的影响

Fig.4 Effects of the grain filling regulators on the gene expression level of key enzymes of sucrose-starch metabolism in inferior spikelets of rice

图5 灌浆充实调节剂对水稻弱势籽粒中Z+ZR和IAA含量的影响

Fig.5 Effects of the grain filling regulators on the contents of Z+ZR and IAA in inferior spikelets of rice

图6 水稻弱势籽粒灌浆速率与miR1432、OsGF14f表达量及IAA含量的相关性

Fig.6 Correlation between the filling rate and the expressions of miR1432, OsGF14f,and the content of IAA in inferior spikelets of rice

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