公主岭霉素诱导对育苗期水稻耐冷性的影响

doi:10.3864/j.issn.0578-1752.2020.11.006

Abstract

Abstract:

【Objective】The objective of this study is to investigate the effects of gongzhulingmycin on rice seedling growth, cold tolerance genes expression and defense enzyme activities based on chilling stress exposure, and to elucidate the change in cold tolerance induced by gongzhulingmycin at the breeding stage of rice. 【Method】Rice variety ‘Jijing88’ was used as the experimental material, the seed germination and seedling growth were investigated under different temperatures and the critical temperature of seed germination was calculated after seed priming by gongzhulingmycin. The solid fermentation product of gongzhulingmycin was powdered and added into the rice seedling substrate before sowing. Taking ‘Jijing88’ as the experimental material, the rice seedlings were subjected to cold stress treatment in simulated cold environment in late spring at the needle appearance stage of rice. Gradually warmed up and sampled continuously within 1-8 d after treatment and sampled every 7 days when plant temperature raised to 28℃. Taking ‘Jihong 6’ as the experimental material, the rice seedlings were raised in the greenhouse and sampled when the seedlings grew to one leaf at a time. The expression of 4 cold tolerance genes (OsNAC6, OsSADMC, OsETR4 and OsZFP151) under the simulated cold environment in late spring and greenhouse seedling environment was analyzed by real-time quantitative PCR (RT-qPCR). The changes of defense enzyme activities such as superoxide dismutase (SOD), catalase (CAT), phenylalanine ammonia-lyase (PAL), polyphenol oxidase (PPO) in greenhouse seedlings before transplanting were detected. 【Result】The gongzhulingmycin could improve seed germination rate and germination index, shorten mean germination time, and its promoting effect on seedling growth was gradually obvious with the decrease of temperature. G-500x showed the best performance, which reduced the critical temperature of rice seed germination by 4.09%, and the subsequent experimental study was carried out based on this concentration. At the condition of low temperature simulation of the late spring cold, the application of gongzhulingmycin significantly increased the expression of OsNAC6, OsSADMC, OsETR4, and rapid response could be achieved to cope with low temperature stress by accelerating the response speed. The control mainly regulated the response speed and expression of OsZFP151 to cope with low temperature stress, but the expression of OsZFP151 was also increased after gongzhulingmycin application compared to the control. After the rice seedlings were subjected to low temperature stress at the needle appearance stage of rice, the expression peaks of OsNAC6, OsSADMC and OsETR4 in the seedlings treated with gongzhulingmycin appeared 1-2 d earlier than that in the control, and were increased by 38.57%, 74.66% and 130.61%, respectively, compared with the control. The maximum expression of OsZFP151 was 2 d later than that of the control, but the maximum expression was 34.91% higher than that of the control. The optimum added weight of the solid fermentation productive powder of gongzhulingmycin was 8 g·m ^-2 in rice seedling substrate in the greenhouse seedling raising. When the seedlings grew to one leaf at a time after the addition of gongzhulingmycin, the expression levels of OsNAC6, OsSADMC, OsETR4 and OsZFP151 in the leaves were higher than those in the control group. The expression of OsNAC6, OsSADMC and OsETR4 was significantly increased. When the added weight was 8 g·m ^-2, the expression level of OsNAC6 was the highest (261.20). When the added weight was 5 g·m ^-2, the expression level of OsSADMC was the highest, which was increased by 126.30% than that of the control. When the added weight was 8 g·m ^-2, the expression level of OsETR4 was the highest, which was increased by 359.81% than that of the control. Gongzhulingmycin could increase the defense enzyme activities of rice seedlings at the four-leaf stage before transplanting. The activities of SOD, POD, PPO and PAL were all increased, especially the SOD and PPO activities increased by 57.18% and 28.53%, respectively. 【Conclusion】Proper application of gongzhulingmycin before sowing can decrease the critical temperature of germination, promote the growth of seedlings, significantly improve the seedling quality, increase the expression levels of cold tolerance genes, improve the response speed to low temperature stress and raise defense enzyme activities to stimulate the cold tolerance in rice seedlings.

Key words: gongzhulingmycin, rice, cold tolerance, gene expression, defense enzyme activity

AN JunXia,ZHAO Yu,ZHANG ZhengKun,SHI HaiPeng,JI DongMing,CAO HongYi,DU Qian,LI QiYun. Induction of Cold Tolerance in Rice at the Breeding Stage by Gongzhulingmycin[J].Scientia Agricultura Sinica, 2020, 53(11): 2195-2206.

Figures/Tables 7

Table 1

Table 2

The effects of gongzhulingmycin on seed germination and seedling growth"

		16℃	18℃	20℃	22℃	24℃	26℃
发芽时间Germination time (d)	CK	8.81±0.04a	9.10±0.06a	6.70±0.02a	5.95±0.01a	5.89±0.02b	3.91±0.01a
	Y-1000X	7.53±0.04 b	8.98±0.09a	6.86±0.04a	5.99±0.02a	6.03±0.01a	3.89±0.00ab
	G-500X	8.21±0.17ab	8.98±0.14a	6.54±0.06b	5.96±0.04a	5.97±0.04ab	3.87±0.01b
	G-1000X	8.75±0.25a	8.98±0.14a	6.54±0.06b	6.06±0.01a	5.99±0.01a	3.92±0.00a
发芽率 Germination rate (%)	CK	71.00±2.00b	78.00±7.51a	91.33±2.91a	97.00±1.00a	88.00±1.15a	96.33±0.33a
	Y-1000X	64.00±2.65b	70.00±3.51a	92.00±3.00a	95.00±0.58a	88.67±1.45a	95.33±0.33a
	G-500X	79.67±1.45a	79.00±3.61a	92.67±0.67a	94.67±0.33a	89.67±0.33a	96.33±0.33a
	G-1000X	69.67±1.33b	77.00±2.31a	91.67±0.89a	94.67±0.33a	85.00±0.58a	96.33±0.33a
发芽指数Germination index	CK	58.65±0.85b	81.70±6.81a	64.78±2.16b	134.87±0.42a	127.47±3.27a	156.93±0.82ab
	Y-1000X	71.33±4.47ab	74.78±0.53b	70.60±2.28b	132.65±1.89a	114.93±1.02b	158.91±0.96ab
	G-500X	81.20±7.64a	83.67±6.90a	84.55±3.08a	133.17±2.66a	121.56±3.88ab	161.91±2.06a
	G-1000X	60.33±7.02b	74.67±6.90b	84.55±3.08a	131.74±1.21a	115.13±0.74b	154.19±0.85b
幼苗株高 Seedling height (cm)	CK	0.93±0.16b	1.72±0.48a	3.10±0.24ab	6.51±0.30a	21.50±0.19a	22.73±0.69a
	Y-1000X	1.52±0.10a	0.98±0.10a	2.48±0.29b	6.34±0.48a	16.96±0.43b	14.46±0.09b
	G-500X	1.25±0.18ab	1.38±0.25a	3.81±0.43a	6.08±0.35a	17.93±1.66b	13.70±0.57bc
	G-1000X	1.25±0.18ab	1.01±0.08a	2.63±0.36b	6.06±0.24a	15.80±0.40b	11.69±0.09c
幼苗鲜重 Seedling fresh weight (g)	CK	0.07±0.01b	0.12±0.01a	0.18±0.01a	0.32±0.02a	0.69±0.03a	0.73±0.031a
	Y-1000X	0.11±0.00a	0.08±0.01b	0.18±0.002a	0.36±0.03a	0.70±0.04a	0.66±0.02b
	G-500X	0.08±0.01b	0.10±0.004ab	0.17±0.01a	0.27±0.02a	0.68±0.02a	0.64±0.02b
	G-1000X	0.08±0.01b	0.08±0.001b	0.17±0.01a	0.27±0.02a	0.59±0.02a	0.63±0.01b
幼苗干重 Seedling dry weight (g)	CK	0.009±0.0003b	0.013±0.001a	0.027±0.001a	0.047±0.003ab	0.100±0.006a	0.074±0.002a
	Y-1000X	0.019±0.004a	0.008±0.001b	0.026±0.001a	0.054±0.003a	0.100±0.006a	0.081±0.002a
	G-500X	0.013±0.001ab	0.013±0.001a	0.029±0.001a	0.046±0.003ab	0.099±0.000a	0.081±0.003a
	G-1000X	0.011±0.001ab	0.008±0.001b	0.027±0.001a	0.042±0.003b	0.083±0.003a	0.081±0.002a

Table 2

Table 3

Fig. 1

Fig. 2

Fig. 3

Fig. 4

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基因 Gene	功能 Function	引物序列 Primer sequence
OsNAC6^[24]	转录因子Transcription factor	F: 5′TCCGCCCGCAAGAGAACAG3′ R: 5′ACCCCCACCATCGGCTTCCT3′
OsSADMC^[2]	S-腺苷甲硫氨酸脱羧酶S-adenosylmethionine decarboylase	F: 5′ATGGTGAACCCTGCTCCT3′ R: 5′GCAGAAGGCAAAGGAAAT3′
OsETR4^[25]	乙烯受体Ethylene receptor	F: 5′GTCTTCCACCTGCTGCTTG3′ R: 5′CCTCTCTTCACCCACATTGC3′
OsZFP151^[26]	C2H2型锌指蛋白C2H2 zinc finger protein	F: 5′GCTCTCCATCAGTTGCTCCT3′ R: 5′CTTCATCATCCCACACCAGA3′
OsActin^[27]	内参基因Internal reference gene	F: 5′CCTGGCAGTATGAAGGTAGTTG3′ R: 5′GAAGCACTTCATGTGGACGAT3′

处理 Treatment	回归方程 Regression equation (y=)	相关系数 Correlation coefficient (r)	信度水平 Reliability level (a)	临界温度 Critical temperature (℃)
CK	10.112x-108.78	0.8381	0.01	10.76
Y-1000X	8.8629x-82.253	0.7864	0.05	9.28
G-500X	9.4620x-97.665	0.7181	0.05	10.32
G-1000X	9.7130x-103.54	0.9333	0.01	10.66

Induction of Cold Tolerance in Rice at the Breeding Stage by Gongzhulingmycin

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