公主岭霉素诱导对育苗期水稻耐冷性的影响

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安俊霞¹^,², 赵宇¹, 张正坤¹, 史海鹏³, 纪东铭⁴, 曹洪翼⁵, 杜茜^,¹, 李启云^,¹

¹吉林省农业科学院植物保护研究所/吉林省农业微生物重点实验室/农业农村部东北作物有害生物综合治理重点实验室,长春 130033

²吉林农业大学植物保护学院,长春 130118

³伊通满族自治县农业技术推广总站,吉林伊通 130700

⁴四平市植物保护站,吉林四平 136000

⁵哈尔滨师范大学生命科学与技术学院,哈尔滨 150080

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

AN JunXia¹^,², ZHAO Yu¹, ZHANG ZhengKun¹, SHI HaiPeng³, JI DongMing⁴, CAO HongYi⁵, DU Qian^,¹, LI QiYun^,¹

¹Institute of Plant Protection, Jilin Academy of Agricultural Sciences/Jilin Key laboratory of Agricultural Microbiology/Key Laboratory of Integrated Pest Management on Crops in Northeast, Ministry of Agriculture and Rural Affairs, Changchun 130033

²College of Plant Protection, Jilin Agricultural University, Changchun 130118

³Agricultural Technology Extension Station of Yitong Manchu Autonomous County, Yitong 130700, Jilin

⁴Plant Protection Station of Siping City, Siping 136000, Jilin

⁵Life Science and Technology Academy, Harbin Normal University, Harbin 150080

通讯作者: 李启云,E-mail： qyli@cjaas.com; 杜茜,E-mail： dqzjk@163.com

责任编辑: 岳梅
收稿日期:2019-10-30接受日期:2019-12-23网络出版日期:2020-06-01

基金资助:

国家重点研发计划.2017YFD0201104
吉林省农业科技创新工程自由创新项目.CXGC2018ZY022
吉林省农业科技创新工程研究生基金.安俊霞 2018

Received:2019-10-30Accepted:2019-12-23Online:2020-06-01
作者简介 About authors
安俊霞,E-mail： 1780120470@qq.com。

摘要
【目的】探讨在低温条件下公主岭霉素对水稻幼苗生长及耐冷相关基因表达、抗逆防御酶活性的影响,明确施用公主岭霉素后育苗期水稻耐冷性的变化。【方法】以‘吉粳88’为试验材料,经公主岭霉素水浸提液浸种引发后播种,调查不同温度下稻种萌发和幼苗生长状况并计算稻种发芽临界温度。将农抗“769”固体发酵物干燥粉碎后添加到水稻育苗基质中,以‘吉粳88’为试验材料,在立针期模拟倒春寒生境对水稻幼苗施加冷胁迫处理,在处理后逐渐缓慢升温并于1—8 d内连续采样,升温至28℃后每隔7 d采样一次;以‘吉宏6号’为试验材料,大棚育苗待水稻幼苗长至一叶一心期采样;采用实时荧光定量PCR（RT-qPCR）技术,分析模拟生境及大棚育苗环境中幼苗叶片耐冷相关基因OsNAC6、OsSADMC、OsETR4、OsZFP151的表达状况,并检测移栽前大棚秧苗叶片中防御酶超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、苯丙氨酸解氨酶（PAL）、多酚氧化酶（PPO）活性的变化。【结果】公主岭霉素能够提高稻种发芽率和发芽指数,缩短发芽时间,其对幼苗生长的促进作用随温度的降低而逐渐明显,G-500X表现最好,使稻种萌发的临界温度降低了4.09%,并以该浓度为基础进行后续的试验研究。光照培养箱低温模拟倒春寒气候条件,低温胁迫下公主岭霉素可调控OsNAC6、OsSADMC、OsETR4表达量的提升,并通过加快响应速度,实现快速反应以应对低温胁迫,对照处理主要通过调控OsZFP151的响应速度和表达量提升应对低温胁迫,但公主岭霉素处理下的水稻幼苗该基因表达量亦得到提升。水稻幼苗在立针期受到低温胁迫后,公主岭霉素处理的幼苗在低温胁迫后OsNAC6、OsSADMC、OsETR4表达量峰值出现时间较对照早1—2 d,且分别比对照提高38.57%、74.66%、130.61%;OsZFP151最高表达量的时间较对照迟2 d,但最高表达量比对照提高了34.91%。大棚育苗,固体发酵物干粉在育苗基质中的最适添加量为8 g·m ^-2,添加后一叶一心期的水稻幼苗叶片中OsNAC6、OsSADMC、OsETR4、OsZFP151的表达量均高于对照,其中OsNAC6、OsSADMC、OsETR4表达量显著提升,OsNAC6在添加量为8 g·m ^-2时表达量最高,为261.20;当固体发酵物干粉添加量为5 g·m ^-2时,OsSADMC表达量最高,比对照提高了126.30%;当固体发酵物干粉添加量为8 g·m ^-2时,OsETR4表达量最高,比对照提高了359.81%。移栽前处于四叶一心期的幼苗,育苗基质中添加公主岭霉素诱导幼苗叶片中SOD、POD、PPO、PAL酶活性均高于对照,其中起主要作用的酶为SOD、PPO,活性分别比对照提高57.18%、28.53%。【结论】公主岭霉素在水稻育苗期施用可降低稻种的发芽临界温度,促进幼苗生长,显著提升幼苗的秧苗素质;公主岭霉素通过提升耐冷相关基因的表达量、加快低温胁迫的响应速度及提高幼苗体内防御酶的活性,促进水稻幼苗耐冷性的增强。
关键词： 公主岭霉素;水稻;耐冷性;基因表达;防御酶活性

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.

Keywords：gongzhulingmycin;rice;cold tolerance;gene expression;defense enzyme activity

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本文引用格式
安俊霞, 赵宇, 张正坤, 史海鹏, 纪东铭, 曹洪翼, 杜茜, 李启云. 公主岭霉素诱导对育苗期水稻耐冷性的影响[J]. 中国农业科学, 2020, 53(11): 2195-2206 doi:10.3864/j.issn.0578-1752.2020.11.006
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 Acricultura Sinica, 2020, 53(11): 2195-2206 doi:10.3864/j.issn.0578-1752.2020.11.006

0 引言

【研究意义】水稻是世界三大粮食作物之一,是我国最主要的粮食作物^[1]。作为喜温作物,其对温度反应敏感,温度在15—20℃时就会对水稻的正常生长产生不利影响^[2]。我国北方地区早春气候异常,低温对植物种子的萌发和幼苗的生长有很大的影响,而种子萌发很大程度上决定了植物的出苗、生长以及收获时稻米的产量和品质。【前人研究进展】提高植物耐冷性比较有效的方法包括采用传统育种培育耐低温品种,如荷兰育种专家培育出抵抗12℃的黄瓜品种,以及我国育种专家选育的耐低温品种‘津优3号’‘津优20号’等^[3];植物经低温处理后,细胞结构和细胞内各种物质将发生一系列形态和生理生化等方面的适应性变化^[4],如细胞膜透性变大^[5],丙二醛（malondialdehyde,MDA）含量升高^[6],超氧化物歧化酶（superoxide dismutase,SOD）、过氧化氢酶（catalase,CAT）、过氧化物酶（peroxidase,POD）、苯丙氨酸解氨酶（phenylalanine ammonia-lyase,PAL）等保护酶的活性升高^[7,8,9],因此通过种子变温处理、低温锻炼等方法可以提高植物耐冷性^[10,11];增施化学肥料、脱落酸（ABA）^[12]和多效唑^[13]等化学试剂,亦可提高植物耐冷能力;另有研究表明,利用空气和植物根系部分不同种类的有益微生物来促进植物生长,可以提高植物对逆境的抵抗能力^[14,15,16],进而克服低温对植物产生的不利影响。不吸水链霉菌公主岭变种（Streptomyces ahygrosscopicus gongzhulingensis n. var.）（农抗“769”）是一株对病原真菌具有广谱抑菌作用的生防链霉菌,其代谢产物被称为公主岭霉素,是一种自然生物合成的混合制剂,具有较好的水溶性,通过液体发酵或固体培养均可获得。研发初期,农抗“769”及公主岭霉素主要应用于玉米、高粱、小麦等作物土传病害的防治^[17]。【本研究切入点】农抗“769”及公主岭霉素可诱导植株体内防御酶活性的变化,从而提高植株的抗病性,并对植物具有促进生长和提升产量、品质的作用^[18,19],然而公主岭霉素诱导对植物应对非生物胁迫的影响尚未见报道。【拟解决的关键问题】通过考察公主岭霉素引发后稻种发芽临界温度、幼苗在低温时的生长状况、幼苗耐冷基因表达及防御酶活性的变化,探究公主岭霉素对水稻耐冷能力的影响,为提升水稻秧苗素质、解决育苗期低温冷害对水稻生产带来损失的问题提供新思路,为应用于水稻育苗期以公主岭霉素为主要成分的生物制剂的研发和施用方式的确定提供数据支持。

1 材料与方法

1.1 供试材料

水稻品种：‘吉粳88’‘吉宏6号’,市售。生物炭无土育苗基质购自沈阳凡宇园艺科技有限公司。芸苔素内酯（Y）市售,EZ-DNA away RNA Min-Preps Kit、AVM第一链cDNA合成试剂盒、2×SG Fast qPCR Master Mix均购自上海生工生物工程有限公司。

农抗“769”固体发酵物干粉：以玉米碴为培养基质,灭菌后接种农抗“769”,28℃充分培养后自然晾晒,干燥并粉碎^[20]。公主岭霉素水浸提液：将农抗“769”固体发酵物按质量体积比1﹕2以纯净水4℃静置浸提24 h,离心取上清液^[20]。

农抗“769”固体发酵物及公主岭霉素水浸提液质量检测：农抗“769”固体发酵充分培养后按质量体积比1﹕2加入无菌水,平板涂布,检测培养物含菌量,达到1×10⁶ cfu/mL以上为充分培养。充分培养后的农抗“769”固体发酵培养物自然晾晒,干燥后粉碎,过100目筛备用。将培养物粉末与水按质量体积比1﹕2混合,制备水浸提液;以玉米大斑病菌（Setosphaeria turcica）为指示菌,通过抑菌圈法检测水浸提液对病原菌的抑制情况,抑菌圈直径达到3.0 cm以上;将符合上述两个指标的培养物用于实验和生产中。

1.2 试验设计

用光照培养箱在实验室模拟水稻遭遇倒春寒的生境,考察公主岭霉素诱导下水稻幼苗的耐冷性。利用光照培养箱种植水稻,生长条件设定为温度28℃、光照12 h·d^-1、相对湿度75%、光照强度5 000 lx,播种后用公主岭霉素（G）水浸提液500倍稀释液（G-500X）将土壤完全润透,以清水润湿的土壤为对照,每个处理设3次重复。待幼苗长至立针期后进行低温胁迫,低温胁迫时保持光照10 h·d^-1、相对湿度75%、光照强度4 000 lx,温度变化过程包括：10℃胁迫1 d,16℃胁迫7 d,恢复28℃培养14 d。从低温胁迫起始日每天采集幼苗叶片,-80℃保存,用于测定幼苗叶片中耐冷相关基因表达量变化。

生产性大棚育苗考察公主岭霉素诱导下水稻幼苗耐冷性试验于2019年4月14日至5月8日在吉林省伊通满族自治县建国村育苗大棚中进行。育苗大棚在育苗期间田间管理及农事操作专人专管,除受气候因素影响外,其他保持不变。棚内生长温度约为21—25℃,相对湿度为50%—90%。实验室模拟环境公主岭霉素最佳使用量为1.5 g·m^-2^[19],本研究中,以实验室模拟环境中的最佳使用量为起点,在田间生产中以添加量差异设置质量梯度,以不添加为对照,分别以1.5、3、5、8、10 g·m^-2的添加量将粉碎的农抗“769”固体发酵物拌入育苗基质中,至幼苗长至一叶一心期,采集幼苗叶片-80℃保存,用于测定幼苗叶片中耐冷相关基因表达量变化。以耐冷相关基因表达量数据为参考,选择基因表达量高的处理,待幼苗长至四叶一心期,移栽前采集叶片,-80℃保存,测定幼苗叶片中防御酶活性的变化。以不添加作为对照,每个处理设3次重复。

1.3 方法

1.3.1 种子萌发及幼苗生长的测定选取大小均匀一致且籽粒饱满的水稻种子,参照前期研究结果设置公主岭霉素水浸提液浓度^[19]。分别以公主岭霉素水浸提液500倍稀释液（G-500X）、1 000倍稀释液（G-1000X）,芸苔素内酯1 000倍稀释液（Y-1000X）及清水浸种引发,25℃浸种48 h,每处理设3次重复,每个重复100粒种子。育苗盘中称取相同质量的基质,将引发后的种子分别播种于育苗盘中,分别在16、18、20、22、24、26℃黑暗条件下萌发^[21],每天统计发芽数（以幼芽长度达到种子长度的1/2,且幼根与种子等长为发芽标准,两者需要同时满足）,每天记录3次。计算发芽率、发芽指数、平均发芽时间、临界温度等。计算公式如下：

（1）G=$\frac{n}{N}\times$ 100%

式中,G：种子发芽率,n：发芽种子数,N：供试种子总数。

（2）GI=Σ Gt/Dt

式中,GI：发芽指数,Gt：逐日发芽种子数,Dt：相应发芽天数。

（3）T=$\frac{\sum{{Gt}/{Dt}\;}}{\sum{Gt}}$

式中,T：平均发芽时间。

发芽临界温度的计算是由发芽指数（y）对发芽温度（x）作回归分析,回归曲线与x轴交点（y=0）即为低温发芽起始。发芽指数与温度的回归关系在0.01和0.05差异水平上经t测验考察回归方程的可靠性^[21,22]。

待水稻发芽后按昼/夜=12 h/12 h生长至第10天,测定株高、幼苗鲜重,并将幼苗在电热恒温干燥箱100℃杀青10 min,80℃烘12 h至恒重,称干重。

1.3.2 水稻幼苗叶片耐冷基因表达量测定采用实时荧光定量PCR（RT-qPCR）技术对幼苗叶片耐冷相关基因进行表达差异分析。以水稻组成型表达基因Actin作为内参基因,以相对定量的方法对目的基因的表达量进行检测。采用2^-??CT法计算相对表达量^[23],每个试验样品进行3次独立的生物学重复。目标基因、基因功能及引物序列见表1。

Table 1
表1
表14个耐冷相关基因RT-qPCR的引物
Table 1Primers of 4 cold tolerance genes for RT-qPCR

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′

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1.3.3 水稻幼苗叶片防御酶活性测定苯丙氨酸解氨酶（PAL）的活性测定采用苯丙氨酸紫外吸收法^[28];多酚氧化酶（PPO）的活性测定采用邻苯二酚-紫外吸收法^[28];超氧化物歧化酶（SOD）的活性测定采用氮蓝四唑自氧化法^[29];过氧化氢酶（CAT）的活性测定采用钼酸铵法^[30]。

1.4 数据处理

采用Excel 2007处理数据及作图,采用DPS15.10高级版进行单因素方差分析,IBM SPSS Statistics 20进行回归分析（Tukey法）。

2 结果

2.1 公主岭霉素引发后水稻在冷胁迫下的生长变化

随种植温度降低,稻种发芽时间增长,发芽率和发芽指数均呈下降趋势,幼苗生长速度、鲜重和干重均降低。公主岭霉素引发后,在稻种发芽至二叶一心生长期前,不同生长温度表现出不同的作用效果。在生长温度较高时,水稻幼苗会出现徒长的现象。如表2所示,24—26℃恒温种植,公主岭霉素处理稻种其作用效果主要表现在墩苗壮苗上,幼苗株高比对照显著降低,G-500X和G-1000X均有较好的效果。22℃恒温种植较适宜水稻的生长,除幼苗干重外各处理间各生长指标均无显著差异,但G-500X和G-1000X幼苗鲜重和干重与对照相比均有所降低,表明幼苗叶片的含水量及鲜嫩程度降低,有助于提高幼苗抵御如病害侵染、温度骤降等逆境的能力。16—20℃低温逆境下,公主岭霉素处理水稻种子可以提高种子发芽率和发芽指数,缩短发芽时间,促进幼苗生长。与26℃相比,种植温度为16℃时CK、Y-1000X、G-500X、G-1000X等不同处理间平均发芽时间分别延迟了4.90、3.64、4.34、4.83 d,发芽率分别降低了25.33%、31.33%、16.66%、26.66%,发芽指数分别降低98.28%、87.58%、80.71%、93.86%,G-500X引发后稻种发芽指数最高。公主岭霉素引发后,16℃低温逆境,G-500X处理稻种各生长指标表现最好,平均发芽时间比对照降低了6.81%,发芽率、发芽指数和株高分别比对照提高了12.21%、38.45%和34.41%。

Table 2
表2
表2公主岭霉素对种子发芽及幼苗生长的影响
Table 2The 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

The data in the table are shown as mean ±SE, different lowercase letters after the data indicate significant differences among different treatments of the same temperature and the same survey index (P<0.05)
表中数据为平均值±标准误,数据后不同小写字母表示同一温度同一调查指标的不同处理间差异显著（P<0.05）

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2.2 公主岭霉素对水稻育苗期耐冷性的影响

2.2.1 公主岭霉素引发对稻种发芽临界温度的影响发芽临界温度愈低,种子发芽时对低温的耐受能力愈强^[21]。本研究中,种子在不同引发条件下,发芽临界温度略有差异,表现为CK>G-1000X>G-500X>Y-1000X（表3）,其中,G-500X浸种引发后的发芽临界温度为10.32℃,比对照降低了4.09%。

Table 3
表3
表3公主岭霉素对种子发芽临界温度的影响
Table 3The effect of gongzhulingmycin on critical temperature of seed germination

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

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2.2.2 公主岭霉素诱导对水稻幼苗耐冷基因表达的影响水稻幼苗在立针期遭遇低温胁迫后,叶片中OsNAC6、OsSADMC、OsETR4及OsZFP151先后对胁迫作出了响应,4个基因最高表达量均高于对照,其中,OsNAC6最高表达量为89.50,而对照仅为64.59,比对照提高38.57%;OsSADMC最高表达量为85.74,比对照提高74.66%;OsETR4最高表达量为84.38,比对照提高130.61%;OsZFP151最高表达量为29.91,比对照提高34.91%。4个基因的表达均呈单峰趋势,主要在胁迫早期表达,施用公主岭霉素加快了幼苗对低温胁迫的响应速度,OsNAC6、OsSADMC和OsETR4均比对照响应速度快,基因表达的峰值出现时间分别在冷胁迫后的1、3和6 d,分别比对照提前2、1和2 d,而对照中OsZFP151最先对胁迫作出了响应（图1）。

图1

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图1模拟倒春寒环境下公主岭霉素对水稻幼苗耐冷基因表达的诱导效果

Fig. 1The induction effect of gongzhulingmycin on cold tolerance genes expression of rice seedlings under the simulated cold environment in late spring

水稻育苗时在育苗土中添加农抗“769”固体发酵物粉末,不同添加量对幼苗耐冷基因表达产生的影响略有不同,但OsNAC6、OsSADMC、OsETR4的相对表达量均比对照高。如图2,OsNAC6相对表达量在添加量为8 g·m^-2时最高,5 g·m^-2其次,表达量分别为261.20、161.29;OsSADMC在添加量5 g·m^-2时最高,8 g·m^-2其次,分别比对照高126.30%、100.10%;OsETR4在添加量8 g·m^-2时最高,5 g·m^-2其次,分别比对照高359.81%、113.73%。OsZFP151的相对表达量与其他3个基因的变化趋势不同,随着添加量的增加,基因表达量先下降后上升,在添加量为8 g·m^-2时比对照高1.46%,差异不显著。水稻育苗期以5—8 g·m^-2农抗“769”固体发酵物拌土较为适宜,对水稻幼苗耐冷性的提升有促进作用。

图2

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图2生产性育苗中公主岭霉素对水稻幼苗耐冷基因表达的诱导效果

Fig. 2The induction effect of gongzhulingmycin on cold tolerance genes expression of rice seedlings in the seedling raising in production

2.2.3 公主岭霉素对水稻叶片防御酶活性的影响水稻育苗时在育苗土中添加农抗“769”固体发酵物粉末,水稻幼苗长至四叶一心期时,幼苗叶片中SOD、POD、PPO、PAL等防御酶活性均较对照显著提升。SOD活性为343.02 U/mg FW,比对照提高了57.18%（图3）;对苯丙烷类代谢途径的影响主要表现在显著提高了PPO的活性,其活性为501.80 U/mg FW,比对照提高了28.53%（图4）。

图3

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图3公主岭霉素对水稻幼苗叶片主要植物氧化酶活性的影响

Fig. 3The effects of gongzhulingmycin on plant oxidase activities of the rice seeding

图4

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图4公主岭霉素对水稻幼苗叶片苯丙烷类代谢关键酶活性的影响

Fig. 4The effects of gongzhulingmycin on activities of key enzymes in phenylpropanoid metabolism of the rice seeding

3 讨论

发芽率、发芽势和发芽指数等指标均是衡量种子发芽质量的关键指标,可以反映种子发芽的速度和整齐度^[31],发芽临界温度与田间苗期耐冷性总选择指数紧密相关,通过室内鉴定可以对田间耐冷反应作出可靠估计,是评估水稻耐冷性有效的鉴定方法^[21]。低温条件下水稻种子发芽率和发芽势降低^[32],本研究中,适宜浓度的公主岭霉素处理稻种对稻种发芽和幼苗生长无不利影响,公主岭霉素通过诱导提高种子发芽率、发芽指数,促进幼苗生长、缩短发芽时间应对低温胁迫,提高水稻幼苗的耐冷性。

在田间生产中,水稻在立针期最易受到冷害的影响,从而降低幼苗的生长势和秧苗素质,导致育苗期立枯病发生,移栽时水稻秧苗素质降低,秧苗田间抗病性差,收获时稻米产量和品质降低,因此立针期水稻秧苗的耐冷性对于水稻生产具有巨大的影响。植物通过响应逆境胁迫的信号传导途径将胁迫信号传递至细胞内,调控基因表达并对逆境胁迫作出适应性反应^[33]。NAC转录因子是植物所特有的转录因子,OsNAC6是细胞核内基因组基因,在植物对逆境胁迫的响应和生长发育的过程中扮演非常重要的角色,在整合和转导非生物和生物逆境胁迫信号过程中也发挥着重要的作用^[24,34],低温、干旱、高盐、脱落酸、茉莉酸、机械损伤和病原菌侵染都会诱导其表达,OsNAC6过表达的植株呈现出较高的抗逆性^[24]。腐胺、亚精胺和精胺等多胺广泛参与植物生长发育和响应环境刺激,是植物适应逆境胁迫的重要调节物质^[35],S-腺苷甲硫氨酸脱羧酶（SADMC）是多胺合成代谢过程中起调控作用的关键酶^[36],作为一种调控因子,主要调控植物体内的腐胺、亚精胺和精胺等多胺的积累量,从而影响植物体内的DNA、RNA和蛋白质生物合成,促进植物的生长发育,影响植物应答环境胁迫,增强植物的抗逆性^[2]。OsETR4是乙烯受体基因,是乙烯信号转导的上游原件,乙烯受体作为乙烯信号传导途径中结合并响应乙烯的初始成分,其通过细胞表面糖蛋白的识别与乙烯结合,改变蛋白质构象、激活或抑制相关响应调控因子或功能基因的表达来完成信号转导^[37]。低温可促进OsETR4的表达,从而提升植株的抗逆性^[25]。C2H2型锌指蛋白主要参与植物生长发育,并且对逆境胁迫的响应有着重要的作用^[38,39,40],OsZFP151受低温诱导后通过调控CBF类基因的表达、影响可溶性糖、脯氨酸含量的增加以及MDA含量的减少来提高水稻耐冷性^[26]。本研究模拟田间倒春寒环境,对立针期幼苗施加低温胁迫,公主岭霉素的施用促进了耐冷基因表达量的提升并加快了幼苗对低温胁迫的响应速度。在生产性育苗过程中,公主岭霉素拌土对水稻幼苗叶片中OsNAC6、OsSADMC及OsETR4表达量提升的促进作用明显,表明公主岭霉素的施用可以提高水稻对低温的耐受能力。

PAL、PPO、SOD、CAT等是植物体内重要的保护酶,其功能包括参与活性氧清除,酚类、木质素和植保素等抗病相关物质的合成,破坏病原菌细胞壁,使植物产生对病原菌的抵抗能力等^{[41,42,43,44]},研究表明,在盐胁迫后,高粱的SOD、POD、CAT和APX（抗坏血酸过氧化物酶）活性均有所提高,且抗性品种比敏感品种增幅大^[45]。在适当的低温胁迫下不同小麦品种的SOD、POD和CAT均有不同程度的提高,抗寒性与SOD、CAT和POD呈极显著正相关^[46]。本研究中,公主岭霉素在播种前施入,幼苗叶片中SOD、CAT、PAL、PPO活性均有提升,其中SOD和PPO活性比对照提升的幅度显著,表明公主岭霉素的施入提升了幼苗应对冷胁迫的能力。

4 结论

（1）适宜浓度的公主岭霉素在低温环境下可以提升稻种的发芽率,促进水稻的幼苗建成,提高水稻的秧苗素质;（2）适宜浓度公主岭霉素诱导,可以提升水稻幼苗叶片中耐冷基因的表达量及对逆境的响应速度;（3）适宜浓度公主岭霉素诱导,可以增强水稻幼苗叶片中防御酶的活性。

综上,在水稻育苗期合理施用公主岭霉素,可以诱导提升水稻幼苗的抗寒能力,增强幼苗苗势,从而降低不利的气候条件对秧苗造成的危害。

参考文献原文顺序
文献年度倒序
文中引用次数倒序
被引期刊影响因子

[1]

朱德峰, 张玉屏, 陈惠哲, 向镜, 张义凯 . 中国水稻高产栽培技术创新与实践
中国农业科学, 2015,48(17):3404-3414.

DOI:10.3864/j.issn.0578-1752.2015.17.008 URL [本文引用: 1]

水稻是中国主要粮食作物,也是单产最高的粮食作物。文章利用中国农业统计年鉴数据和国内外文献资料,分析了中国水稻高产栽培技术创新与实践的成功经验、存在问题和稻作技术发展趋势。探讨了中国的水稻生产经历4个阶段的特点,良种良法配套对水稻增产的贡献,不同阶段创新的稻作技术；简述了全球水稻栽培技术发展的特点；回顾了中国矮秆品种、杂交稻及超级稻品种更替及其配套栽培技术创新；剖析了水稻高产栽培存在的新问题；对水稻高产栽培技术的未来发展进行了展望。创新与品种生育特性配套的水稻种植制度、生产模式和环境协调的栽培技术,发挥品种产量潜力和应用,可以实现增产增效,促进水稻产业发展,改善生态环境和提高资源利用效益。

ZHU D

, ZHANG Y

, CHEN H

, XIANG

, ZHANG Y

. Innovation and practice of high-yield rice cultivation technology in China
Scientia Agricultura Sinica, 2015,48(17):3404-3414. (in Chinese)

DOI:10.3864/j.issn.0578-1752.2015.17.008 URL [本文引用: 1]

胡晓晨, 张婷, 杨圣, 秦巧, 石英尧, 王文生, 傅彬英 . 水稻冷胁迫响应基因OsSADMC功能标记的开发和利用
中国水稻科学, 2015,29(5):475-480.

DOI:10.3969/j.issn.1001-7216.2015.05.004 URL [本文引用: 3]

通过对95份水稻种质资源进行苗期耐冷性鉴定，筛选出10份苗期耐冷（存活率≥60%）和9份冷敏感的材料（存活率为零），并对水稻冷胁迫响应基因OsSADMC进行了表达分析，发现对照和冷胁迫处理条件下该基因在耐冷品种中的表达量都显著高于冷敏感品种。对耐冷材料丽江新团黑谷（LTH）和冷敏感材料IR29的基因编码区域进行了克隆和测序分析，在两个品种间鉴定了10个SNP位点，其中第749碱基处存在的一个碱基变异（C突变为T），从而使丝氨酸变为亮氨酸。根据其中一个SNP位点设计了OsSADMC的功能标记SADMCCAPS1，该标记可以准确地鉴定出19份耐冷性不同水稻种质资源OsSADMC的基因型和耐冷性。 

HU X

, ZHANG

, YANG

, QIN

, SHI Y

, WANG W

, FU B

. Development and application of a functional marker for the cold stress responsive gene
OsSADMC. Chinese Journal of Rice Science, 2015,29(5):475-480. (in Chinese)

DOI:10.3969/j.issn.1001-7216.2015.05.004 URL [本文引用: 3]

马俊 . 丛枝菌根真菌对黄瓜幼苗低温胁迫的缓解效应及其调控机理
[D]. 杨凌: 西北农林科技大学, 2016.

[本文引用: 1]

. Alleviative effects and its mechanism of exogenous arbuscular mycorrhizal fungi (AMF) on cucumber seedlings under cold stress
[D]. Yangling: Northwest A&F University, 2016. ( in Chinese)

[本文引用: 1]

[4]

乌凤章, 王贺新, 徐国辉, 张自川 . 木本植物低温胁迫生理及分子机制研究进展
林业科学, 2015,51(7):116-128.

DOI:10.11707/j.1001-7488.20150713 URL [本文引用: 1]

低温胁迫限制了许多野生植物和作物的地理分布,降低了生产率。植物在低温胁迫下生存能力差别很大,来源于热带和亚热带的冷敏感植物在温度高于冰点的低温条件下会发生不可逆的伤害,而温带地区的植物能够承受极端的冰冻条件。近年来有关冷敏感植物和温带植物低温胁迫下生理生化响应特征,转录组学、蛋白质组学和代谢组学分析取得了较快进展,从不同层次阐明了木本植物调节低温反应和抗冷(冻)性的生理和分子机制,为利用抗冷(冻)相关基因进行林木抗(冷)冻分子育种提供了重要参考。冷敏感树木在1~10℃的低温胁迫下,会出现水分状况、矿质营养、光合作用、呼吸作用和新陈代谢等生理过程的紊乱,造成冷害甚至死亡。钙信号途径是低温应答过程中重要的信号转导途径。ABA通过ABA依赖的转录因子的转录激活参与植物胁迫响应基因的调节。低温响应转录调控分为CBF途径和非CBF途径。CBF调控下游基因的表达过程: 植物细胞膜相关受体首先感知环境信号并传送到细胞核,通过Ca²⁺和MAPKs等感知和传导信号,诱导转录因子CBF的表达,从而激活冷响应基因的启动子,触发冷响应机制(基因激活),转录的mRNA被翻译成不同的蛋白质。这些基因产物参与改变膜质成分组成、抗氧化酶活性、渗透物质含量等生理过程,从而提高抗冷性。另外,CBF基因表达也受到上游一些转录因子的调节,包括钙离子信号途径和ICE1-CBF-寒冷响应途径。树木抗冷锻炼和抗冻锻炼的机制与代谢途径有许多相似之处,但后者可能更复杂,因为它需要面对极端的低温和不正常的温度波动。抗冻锻炼期间低温诱导的抗冻蛋白具有较强的抑制冰晶重结晶的活性; 脱水蛋白和胚胎后期丰富蛋白(LEA)通过渗透调节保护细胞; 热休克蛋白(HSPs)调节蛋白质的折叠和运输,恢复钝化酶的活性; 抗氧化酶系统清除氧自由基和过氧化氢; 早期光诱导蛋白参与高光胁迫适应过程,这些蛋白在树木抗冻机制中具有重要作用。在树木抗冷(冻)领域,未来应加强对控制抗冷(冻)生理变化的转录因子及关键功能基因的全面解析, 深入探讨光、温度等环境信号诱导抗冻性形成机制, 使用蛋白质组学方法与其他技术相结合阐明低温胁迫下重要蛋白质表达机制和功能,完整地揭示木本植物在低温逆境下的生存机制。

WU F

, WANG H

, XU G

, ZHANG Z

. Research progress on the physiological and molecular mechanisms of woody plants under low temperature stress
Scientia Silvae Sinicae, 2015,51(7):116-128. (in Chinese)

DOI:10.11707/j.1001-7488.20150713 URL [本文引用: 1]

SHEIKH

, NOH

, SEONG M

, HJUNG G

, KIM J

. Consequences of chilling stress on watermelon [Citrullus lanatus (Thunb.) Matsum. and Nakai] germplasm lines at seedling stage
Horticulture, Environment, and Biotechnology, 2015,56(1):79-88.

DOI:10.1007/s13580-015-0174-2 URL [本文引用: 1]

[6]

HAWRYLAK-NOWAK

, MATRASZEK

, SZYMA?SKA

. Selenium modifies the effect of short-term chilling stress on cucumber plants
iological Trace Element Research, 2010,138(1/3):307-315.

[本文引用: 1]

[7]

项洪涛, 齐德强, 李琬, 郑殿峰, 王月溪, 王彤彤, 王立志, 曾宪楠, 杨纯杰, 周行, 赵海东 . 低温胁迫下外源ABA对开花期水稻叶鞘激素含量及抗寒生理的影响
草业学报, 2019,28(4):81-94.

[本文引用: 1]

XIANG H

, QI D

, LI

, ZHENG D

, WANG Y

, WANG T

, WANG L

, ZHENG X

, YANG C

, ZHOU

, ZHAO H

. Effect of exogenous ABA on the endogenous hormone levels and physiology of chilling resistance in the leaf sheath of rice at the flowering stage under low temperature stress
Acta Prataculturae Sinica, 2019,28(4):81-94. (in Chinese)

[本文引用: 1]

[8]

XIANG H

, WANG T

, ZHENG D

, WANG L

, FENG Y

, LUO

, LI

, LI Z

, MENG

, LI

, WANG L

, YANG C

. ABA pretreatment enhances the chilling tolerance of a chilling-sensitive rice cultivar
Brazilian Journal of Botany, 2017,40(4):853-860.

DOI:10.1007/s40415-017-0409-9 URL [本文引用: 1]

[9]

董春娟, 李亮, 曹宁, 尚庆茂, 张志刚 . 苯丙氨酸解氨酶在诱导黄瓜幼苗抗寒性中的作用
应用生态学报, 2015,26(7):2041-2049.

URL [本文引用: 1]

为了探讨苯丙氨酸解氨酶(PAL)在诱导黄瓜幼苗抗寒性中的作用，采用喷施特异抑制剂(AOPP)的方法控制PAL活性，测定幼苗抗寒性的变化.结果表明：低温可以诱导黄瓜幼苗叶片中PAL的基因表达和活性升高；喷施AOPP显著抑制了叶片中PAL活性，减少了酚类和类黄酮物质的积累.低温对黄瓜幼苗造成显著伤害，AOPP预处理加剧了低温对幼苗的损伤，幼苗抗寒性降低.与对照相比，幼苗叶片中相对电解质渗漏率和丙二醛(MDA)含量显著升高，PSII最大光化学效率(F_v/F_m)降低，光化学猝灭参数Y(NO)升高，胁迫相关基因(PR1-1a、COR47、P5CS、HSP70)的诱导表达受到抑制.低温导致黄瓜幼苗叶片中H₂O₂积累，还原型抗坏血酸(AsA)含量降低，脱氢抗坏血酸(DHA)含量升高，AsA∶DHA减小；喷施AOPP的幼苗中抗氧化酶(过氧化氢酶CAT、抗坏血酸过氧化物酶APX)活性显著低于对照，H₂O₂过量积累，AsA∶DHA更低.施用H₂O₂清除剂可以有效缓解喷施AOPP引起的低温损伤加剧，而施用CAT抑制剂的幼苗对低温胁迫更敏感.表明低温诱导了PAL活性升高，促进了苯丙烷类次生代谢产物的合成，提高了胞内抗氧化酶活性，可有效清除活性氧分子，维持AsA氧化还原状态，缓解低温引起的光损伤和氧化损伤.

DONG C

, LI

, CAO

, SHANG Q

, ZHANG Z

. Roles of phenylalanine ammonia-lyase in low temperature tolerance in cucumber seedlings
Chinese Journal of Applied Ecology, 2015,26(7):2041-2049. (in Chinese)

URL [本文引用: 1]

JANDA

, MAJLáTH

, SZALAI

, . Interaction of temperature and light in the development of freezing tolerance in plants
Journal of Plant Growth Regulation, 2014,33(2):460-469.

DOI:10.1007/s00344-013-9381-1 URL [本文引用: 1]

Freezing tolerance is the result of a wide range of physical and biochemical processes, such as the induction of antifreeze proteins, changes in membrane composition, the accumulation of osmoprotectants, and changes in the redox status, which allow plants to function at low temperatures. Even in frost-tolerant species, a certain period of growth at low but nonfreezing temperatures, known as frost or cold hardening, is required for the development of a high level of frost hardiness. It has long been known that frost hardening at low temperature under low light intensity is much less effective than under normal light conditions; it has also been shown that elevated light intensity at normal temperatures may partly replace the cold-hardening period. Earlier results indicated that cold acclimation reflects a response to a chloroplastic redox signal while the effects of excitation pressure extend beyond photosynthetic acclimation, influencing plant morphology and the expression of certain nuclear genes involved in cold acclimation. Recent results have shown that not only are parameters closely linked to the photosynthetic electron transport processes affected by light during hardening at low temperature, but light may also have an influence on the expression level of several other cold-related genes; several cold-acclimation processes can function efficiently only in the presence of light. The present review provides an overview of mechanisms that may explain how light improves the freezing tolerance of plants during the cold-hardening period.

[11]

CHEN J

, TIAN Q

, PANG

, JIANG L

, WU R

, XIA X

, YIN W

. Deep-sequencing transcriptome analysis of low temperature perception in a desert tree,
Populus euphratica. BMC Genomics, 2014,15:326.

[本文引用: 1]

[12]

CUEVAS J

, LóPEZ-COBOLLO

, ALCáZAR

, ZARZA

, KONCZ

, ALTABELLA

, SALINAS

, TIBURCIO A

, FERRANDO

. Putrescine is involved in Arabidopsis freezing tolerance and cold acclimation by regulating abscisic acid levels in response to low temperature
Plant Physiology, 2008,148(2):1094-1105.

DOI:10.1104/pp.108.122945 URL [本文引用: 1]

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徐秋曼, 陈宏, 高虹, 程景胜 . 多效唑提高水稻幼苗抗低温能力的机理初探
西北植物学报, 2002,22(5):1236-1241.

[本文引用: 1]

XU Q

, CHEN

, GAO

, CHENG J

. The biochemical mechanism study on improvement of PP₃₃₃ on the cold resistance of the rice seedlings
Acta Botanica Boreali-Occidentalia Sinica, 2002,22(5):1236-1241. (in Chinese)

[本文引用: 1]

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刘润进, 唐明, 陈应龙 . 菌根真菌与植物抗逆性研究进展
菌物研究, 2017,15(1):70-88.

[本文引用: 1]

LIU R

, TANG

, CHEN Y

. Recent advances in the study of mycorrhizal fungi and stress resistance of plants
Journal of Fungal Research, 2017,15(1):70-88. (in Chinese)

[本文引用: 1]

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THEOCHARIS

, CLEMéNT

BARKA E

. Physiological and molecular changes in plants grown at low temperatures
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GARG

, SINGLA

. Naringenin- and Funneliformis mosseae- mediated alterations in redox state synchronize antioxidant network to alleviate oxidative stress in Cicer arietinum L. genotypes under salt stress
Journal of Plant Growth Regulation, 2015,34(3):595-610.

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隋丽, 徐文静, 杜茜, 陈光, 董英山, 李启云 . 放线菌769发酵液对水稻体内主要防御酶活性的影响
吉林农业大学学报, 2009,31(4):382-384, 389.

URL [本文引用: 1]

用放线菌769发酵液处理水稻植株,测定水稻叶片中主要防御酶活性的变化,初步从酶学水平探讨了放线菌769发酵液对水稻的抗性诱导机制。结果表明769菌株发酵液处理水稻可引起水稻体内防御酶发生明显变化,可抑制过氧化氢酶(CAT)的活性,提高过氧化物酶(POD)和苯丙氨酸解氨酶(PAL)活性。以发酵液原液的作用效果最为显著。

SUI

, XU W

, DU

, CHEN

, DONG Y

, LI Q

. Effect of actinomycetes 769 fermentation products on main defense enzyme activity of rice
Journal of Jilin Agricultural University, 2009,31(4):382-384, 389. (in Chinese)

URL [本文引用: 1]

张振鲁, 隋丽, 张佳诗, 李启云, 王金刚, 汪洋洲, 盛岩, 杜茜 . 链霉菌769诱导对百日草抗病性的影响
中国植保导刊, 2013,33(12):14-17.

[本文引用: 1]

ZHANG Z

, SUI

, ZHANG J

, LI Q

, WANG J

, WANG Y

, SHENG

, DU

. Inducing effect of Streptomyces gongzhulingensis 769 on systemic resistance of Zinnia elegans
China Plant Protection, 2013,33(12):14-17. (in Chinese)

[本文引用: 1]

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安俊霞, 李晓光, 汪洋洲, 张正坤, 马嵩岳, 张静, 杨会营, 文松, 杜茜, 李启云 . 公主岭霉素在水稻育秧期应用技术研究
东北农业科学, 2019,44(2):28-33.

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, WANG Y

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, MA S

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, LI Q

. High efficiency application technology of gongzhulingmycin during rice seedling stage
Journal of Northeast Agricultural Sciences, 2019,44(2):28-33. (in Chinese)

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吉林省农科院植保所. 代汞拌种剂农抗769
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Institute of Plant Protection, Jilin Academy of Agricultural Sciences. Mercury seed dressing with Streptomyces gongzhulingensis n. var.
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吴江 . 吉林地区甜玉米种子低温发芽临界温度的研究
种子, 2018,37(3):96-97, 119.

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. Study on the critical temperature of germination of sweet corn seed in Jilin region
Seed, 2018,37(3):96-97, 119. (in Chinese)

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[22]

曾霞, 王彦荣, 胡小文 . 垂穗披碱草种子的萌发适宜温度及温度阈值
草业科学, 2011,28(6):988-992.

URL [本文引用: 1]

以采集于甘肃省和青海省等地的8份垂穗披碱草(Elymus nutans)种质为材料，采用纸上发芽法在5、10、15、20、25、30、35和40 ℃ 8个恒温条件下，研究种子的萌发适宜温度和温度阈值。结果表明，所有供试垂穗披碱草种子在5～35 ℃条件下可萌发，在40 ℃不萌发；以20、25和30 ℃发芽最快，发芽率在3 d内可达最大值，根据种子萌发温度模型计算得出，供试垂穗披碱草种子萌发的最低温度(Tb)、最适温度(To)和最高温度(Tc)的平均值分别为4、24和38 ℃，种质间无显著差异(P＞0.05)。建议垂穗披碱草种子的适宜检验温度为20、25和30 ℃恒温，初次和末次的适宜统计时间分别为第3天和第11天。

ZENG

, WANG Y

, HU X

. Research on seed germination optimum temperature and temperature threshold of Elymus nutans
Pratacultural Science, 2011,28(6):988-992. (in Chinese)

URL [本文引用: 1]

LIVAK

, SCHMITTGEN T

. Analysis of relative gene expression data using real-time quantitative PCR and the 2 ^-ΔΔCT method
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LEE D

, CHUNG P

, JEONG J

, JANG

, BANG S

, JUNG

, KIM Y

, HA S

, CHOI Y

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. The rice OsNAC6 transcription factor orchestrates multiple molecular mechanisms involving root structural adaptions and nicotianamine biosynthesis for drought tolerance
Plant Biotechnology Journal, 2017,15(6):754-764.

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YAU C

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, YIP W

. Differential expression of three genes encoding an ethylene receptor in rice during development, and in response to indole-3-acetic acid and silver ions
Journal of Experimental Botany, 2004,55(397):547-556.

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黄凯 . 水稻C2H2型转录因子OsZFP151在低温胁迫中的功能研究
[D]. 长春: 吉林大学, 2017.

[本文引用: 2]

HUANG

. Functional analysis of C2H2 transcription factor OsZFP151 in rice (Oryza sativa L.) under low temperature stress
[D]. Changchun: Jilin University, 2017. ( in Chinese)

[本文引用: 2]

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关可兴 . 水稻耐冷性相关转录因子OsbZIP32的功能分析
[D]. 长春: 吉林大学, 2015.

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GUAN K

. Functional analysis of transcriptional factor, OsbZIP32 for cold tolerance in rice
[D]. Changchun: Jilin University, 2015. ( in Chinese)

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麦麦提艾力·热合曼, 海利力·库尔班, 郭立华, 邱德文 . 灰霉菌激活蛋白诱导抗病相关的酶活性提高番茄抗病性
中国生物防治学报, 2014,30(6):780-786.

URL [本文引用: 2]

灰霉菌中提取纯化16 kD的激活蛋白.番茄种子以20 mmol/L的HEPES缓冲液(pH 8.0)为对照,用2.5、5、10和20 μg/mL的灰霉菌激活蛋白处理36 h,播种第45 d接种灰霉病,接种第7、14和21 d观察发病率;并用10 μg/mL的激活蛋白溶液喷洒番茄幼苗第6、12、24、48、72和120 h取样测定苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)和多酚氧化酶(PPO)活性.结果显示,10 μg/mL激活蛋白处理番茄种子时,番茄抗病性最强,诱抗效果为48%～54%;番茄幼苗喷洒激活蛋白处理第24 h,PAL活性达峰值、比对照组提高54%,处理第72 h,POD和PPO活性达峰值、分别比对照组提高106%、122%.灰霉菌激活蛋白通过诱导抗病性相关酶的活性,提高番茄的抗病性,并且在最适浓度下诱导效果最佳.

RAHMAN

, KURBAN

, GUO L

, QIU D

. Botrytis cinerea activator protein induce resistance-related enzyme activities and enhancement of disease resistance in tomato plants
Chinese Journal of Biological Control, 2014,30(6):780-786. (in Chinese)

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原向阳, 郭平毅, 张丽光, 王鑫, 赵锐, 郭秀, 宋喜娥 . 干旱胁迫下草甘膦对抗草甘膦大豆幼苗保护酶活性及脂质过氧化作用的影响
中国农业科学, 2010,43(4):698-705.

URL [本文引用: 1]

【目的】为探明干旱胁迫（5 d）及旱后复水条件下不同草甘膦剂量对抗草甘膦大豆（RR1）幼苗保护酶活性及脂质过氧化作用的影响。【方法】采用盆栽试验,在大豆的第三复叶期进行水分胁迫和草甘膦处理。【结果】（1）正常水分条件下,草甘膦增加了RR1的超氧化物歧化酶（SOD）、过氧化物酶（POD）、过氧化氢酶（CAT）活性,以及丙二醛（MDA）含量和相对电导率（EL）,且随剂量的增加和处理5 d内时间的延长而升高;草甘膦处理17 d后,各指标均有所下降。（2）干旱条件下,较低剂量的草甘膦处理使RR1的SOD、POD、CAT活性随胁迫时间的延长而升高,＞0.92 kg?hm-2处理的各保护酶活性随胁迫时间的延长呈先增加后降低趋势;然而,各剂量处理的MDA含量和EL均在胁迫第5天上升到最大;各指标在复水12 d后均有所下降。（3）干旱条件下草甘膦处理的SOD、POD、CAT活性以及MDA含量和EL均高于正常水分条件下草甘膦处理。【结论】正常水分条件下,草甘膦对RR1幼苗造成的伤害可以经过一段时间的生长发育有所缓解;而干旱胁迫加剧了草甘膦对RR1幼苗伤害的原因是活性氧代谢失衡,保护酶系的活性发生变化,质膜过氧化程度加大,短期干旱胁迫后复水可提高细胞膜的抗干旱能力或适应胁迫的能力。

YUAN X

, GUO P

, ZHANG L

, WANG

, ZHAO

, GUO

, SONG X

. Glyphosate and post-drought rewatering on protective enzyme activities and membrane lipid peroxidation in leaves of glyphosate-resistant soybean [glycine max (L.) Merr.] seedlings
Scientia Agricultura Sinica, 2010,43(4):698-705. (in Chinese)

URL [本文引用: 1]

郭彩明, 陈宗明, 陈春丽 . 精氨酸脱羧酶基因AtADC2通过调节超氧化物歧化酶和过氧化氢酶活性增强拟南芥耐盐性
植物生理学报, 2015,51(7):1067-1074.

[本文引用: 1]

GUO C

, CHEN Z

, CHEN C

. AtADC2 enhances salt tolerance through regulating activities of superoxide dismutase and catalase in Arabidopsis thaliana
Plant Physiology Journal, 2015,51(7):1067-1074. (in Chinese)

[本文引用: 1]

[31]

KUMAR

, VERMA S

, SINGH H

. Effect of temperature on seed germination parameters in Kalmegh (Andrographis paniculata Wall.ex Nees.)
Industrial Crops and Products, 2011,34(1):1241-1244.

DOI:10.1016/j.indcrop.2011.04.008 URL [本文引用: 1]

Kalmegh (Andrographis paniculata Wall. ex Nees) is a medicinal herb of tropical south east Asia and native to India and Sri Lanka. It has been used herb for liver ailment in all prevailing systems of medicine viz. Ayurvedic, Unani, Homeopathic and modern throughout most of the south East Asian countries viz. China, India, Sri Lanka, Indonesia, Thailand, Burma, and Vietnam, etc. Commercially cultivation of Kalmegh is done through seeds. Optimal germination potential, temperature and first as well as final count day are three primary parameters for developing the seed quality standards. Germination of Kalmegh variety 'CIM-Megha' was carried out at six constant temperatures at an interval of every '5 -C' from '15 to 40 C' temperatures coupled with 16h light and 8h dark photo period. The temperature of '25 degrees C' was found optimally suitable with '94.6' and '23.6' percentage of germination and germination energy, respectively, while the temperature at '40 degrees C' was deleterious with no germination. Significant decrease in percentage of germination and germination energy was observed at '15 '20 degrees C', '30 and '35 degrees C' of temperature in comparison to '25 degrees C'. The study further revealed that days 5-6 and days 7-9 after seed sowing were the ideal for first and final count, respectively for seed germination of Kalmegh. (C) 2011 Elsevier B.V.

[32]

刘雪梅, 尚庆茂, 张志刚 . 辣椒不同品种种子萌芽期耐低温性及评价方法研究
中国生态农业学报, 2010,18(3):521-527.

URL [本文引用: 1]

以108个辣椒品种为试材, 测定了低温(18 ℃)下种子的相对发芽势、相对发芽率、相对胚根长、相对发芽指数和相对活力指数, 并采用极点排序法对辣椒不同品种的耐低温性进行综合评价。结果表明: 辣椒不同品种种子萌芽期的耐低温性不同, 相对发芽率、相对发芽指数、相对胚根长和相对活力指数4项指标在不同品种间差异达显著水平(P<0.05)。选用上述4个指标, 采用极点排序法计算得到各品种的综合评价得分, 根据综合评价得分将108个辣椒品种分为10个耐低温品种, 9个低温敏感品种, 其余为中度耐低温品种。辣椒种子相对发芽势、相对胚根长与综合评价得分的相关性达到显著水平; 相对发芽率、相对发芽指数、相对活力指数与综合评价得分的相关性达极显著水平, 且相对活力指数与综合评价得分的相关性最高(r=0.899 9)。通过测定低温下辣椒品种萌芽期相关性状指标, 采用极点排序法可对辣椒种子萌芽期耐低温性进行快速、准确的评价。

LIU X

, SHANG Q

, ZHANG Z

. Low-temperature tolerance of pepper at germination stage and its evaluation method
Chinese Journal of Eco-Agriculture, 2010,18(3):521-527. (in Chinese)

URL [本文引用: 1]

郭仰东, 张磊, 李双桃, 曹芸运, 齐传东, 王晋芳 . 蔬菜作物应答非生物逆境胁迫的分子生物学研究进展
中国农业科学, 2018,51(6):1167-1181.

DOI:10.3864/j.issn.0578-1752.2018.06.015 URL [本文引用: 1]

蔬菜作为重要的经济作物,近年来的种植面积、产量及需求都在不断增加。蔬菜作物在生长和发育过程中经常受到非生物逆境（包括干旱、盐、极端温度及重金属胁迫等）的侵害,影响其产量及品质。近十年来,国内外关于蔬菜应答非生物逆境胁迫的分子生物学研究领域取得了一定的进展。在应答干旱胁胁迫方面,DREB、WRKY、NAC、bHLH及bZIP等转录因子受干旱信号诱导,调节下游抗旱基因的表达,从而提高蔬菜作物抗旱能力。同时,水分运输相关功能基因（PIP、TIP）、E3连接酶SIZ1及脱水蛋白DHN也被报道受干旱诱导,并通过调节水势、渗透势及ROS积累抵御干旱胁迫。在抵御盐胁迫方面,SOS途径至关重要。SlSOS2能够通过调节SlSOS1和Na+/H+逆向转运蛋白LeNHX2/4的表达维持离子平衡和调节植物器官中Na+的分配。蔬菜抗盐研究中NAC、ERF、MYB等转录因子响应盐胁迫并激活抗逆相关基因表达,从而提高蔬菜作物抗盐能力。此外蔬菜植物大量合成渗透调节物质是其抵御盐胁迫的常见方式。吡咯啉-5-羧酸合成酶PvP5CS和tomPRO2、脯氨酸脱氢酶BoiProDH等在盐胁迫下能提高脯氨酸的含量；过表达甜菜碱醛脱氢酶SlBADH能提高番茄中甜菜碱含量。在高温胁迫响应过程中,HSFs位于调控网络的核心位置,可调控包括HSPs在内的一系列抗逆基因的表达,番茄中热激转录因子SlHSFs相互之间形成复合体调控下游SlHSPs的表达而应答高温逆境。在低温胁迫中,CBFs/EREBs位于调控网络的核心位置,并受ICE1调控；LEA及HSPs蛋白在低温下能够防止细胞中蛋白质变性并维持细胞膜流动性。蔬菜应答重金属胁迫主要依靠体内隔离和体内外螯合机制。在蔬菜应答非生物逆境的过程中,ABA作为信号受体起到至关重要的作用。蔬菜中NAC、MYB、HSF等转录因子则受ABA信号诱导,应答非生物逆境,进而提高活性氧清除能力,合成更多抗逆物质,从而抵御非生物逆境的侵害。

GUO Y

, ZHANG

, LI S

, CAO Y

, QI C

, WANG J

. Progresses in research on molecular biology of abiotic stress responses in vegetable crops
Scientia Agricultura Sinica, 2018,51(6):1167-1181. (in Chinese)

DOI:10.3864/j.issn.0578-1752.2018.06.015 URL [本文引用: 1]

NAKASHIMA

, TRAN L

, VAN NGUYEN

, FUJITA

, MARUYAMA

, TODAKA

, ITO

, HAYASHI

, SHINOZAKI

, YAMAGUCHI-SHINOZAKI

. Functional analysis of a NAC-type transcription factor OsNAC6 involved in abiotic and biotic stress-responsive gene expression in rice
The Plant Journal, 2007,51(4):617-630.

DOI:10.1111/j.1365-313X.2007.03168.x URL [本文引用: 1]

[35]

KUSANO

, BERBERICH

, TATEDA

, TAKAHASHI

. Polyamines: Essential factors for growth and survival
Planta, 2008,228(3):367-381.

DOI:10.1007/s00425-008-0772-7 URL [本文引用: 1]

Polyamines are low molecular weight, aliphatic polycations found in the cells of all living organisms. Due to their positive charges, polyamines bind to macromolecules such as DNA, RNA, and proteins. They are involved in diverse processes, including regulation of gene expression, translation, cell proliferation, modulation of cell signalling, and membrane stabilization. They also modulate the activities of certain sets of ion channels. Because of these multifaceted functions, the homeostasis of polyamines is crucial and is ensured through regulation of biosynthesis, catabolism, and transport. Through isolation of the genes involved in plant polyamine biosynthesis and loss-of-function experiments on the corresponding genes, their essentiality for growth is reconfirmed. Polyamines are also involved in stress responses and diseases in plants, indicating their importance for plant survival. This review summarizes the recent advances in polyamine research in the field of plant science compared with the knowledge obtained in microorganisms and animal systems.

[36]

ROYA

, WU

. Overexpression of S-adenosylmethionine decarboxylase gene in rice increases polyamine level and enhances sodium chloride-stress tolerance
Plant Science, 2002,163(5):987-992.

DOI:10.1016/S0168-9452(02)00272-8 URL [本文引用: 1]

[37]

张楠楠, 薛冬, 崔晓霞, 赵晋铭, 郭娜, 王海棠, 邢邯 . 大豆组成型三重反应基因GmCTR1的克隆与功能分析
中国农业科学, 2017,50(16):3082-3091.

DOI:10.3864/j.issn.0578-1752.2017.16.003 URL [本文引用: 1]

【目的】植物激素乙烯参与植物的生长发育以及生物与非生物胁迫过程。组成型三重反应基因CTR1作为乙烯受体下游一个负调控因子,结合乙烯受体共同参与乙烯的信号转导途径。本研究通过对大豆（Glycine max）的组成型三重反应基因CTR1进行克隆和诱导表达分析,以及在大豆发状根中过表达该基因后进行疫霉根腐病的抗性分析,探究CTR1在大豆与疫霉菌互作过程中的功能。【方法】利用同源克隆的方法,以拟南芥的AtCTR1序列为探针,在大豆基因组数据库中搜索同源性最高的基因即为大豆的GmCTR1（Glyma.13G151100）,并从Williams82中将GmCTR1克隆出来。对GmCTR1进行多重序列比对、系统进化分析和疫霉菌的诱导表达分析。构建植物过表达载体pBinGFP2: GmCTR1。应用发根农杆菌介导的遗传转化方法获得大豆发状根,经GFP荧光筛选得到大豆过表达GmCTR1的阳性发状根和转入空质粒的阴性对照发状根。对过表达GmCTR1发状根和阴性对照发状根侵染疫霉菌后,检测病斑长度、疫霉生物量和抗性相关基因的表达。【结果】根据同源序列比对结果,将与拟南芥AtCTR1（AT5G03730）同源性最高（75.3%）的大豆基因Glyma.13G151100命名为GmCTR1,从Williams82中克隆得到GmCTR1的CDS序列。该基因的CDS序列全长2 511 bp,编码836个氨基酸的丝氨酸/苏氨酸蛋白激酶,蛋白质量为92.35 kD,等电点为6.51。多重序列比对结果显示,GmCTR1氨基酸具有CTR1同源蛋白的典型结构域和关键特征。构建系统发育树进行进化分析发现,GmCTR1与菜豆（Phaseolus vulgaris）、蒺藜苜蓿（Medicago truncatula）的CTR1亲缘关系十分相近,在同一个分支上,且与菜豆的亲缘关系最近。荧光定量PCR结果表明,在大豆疫霉菌P6497侵染后,GmCTR1受诱导逐渐上调表达,在侵染48 h后表达水平达到最高,随后表达量降低。利用酶切连接方法将GmCTR1的CDS序列构建到植物过表达载体pBinGFP2中,PCR和酶切验证获得了重组质粒pBinGFP2: GmCTR1。对发状根接种疫霉菌后发现,过表达GmCTR1减弱了对疫霉根腐病的抗性,疫霉菌侵染36 h后过表达发状根与对照相比病斑长度显著增长。过表达GmCTR1发状根中疫霉菌的生物量与对照相比增加,与大豆抗病反应相关基因的表达水平显著降低。【结论】GmCTR1在大豆与疫霉菌的互作过程中发挥一定的负调控作用。

ZHANG N

, XUE

, CUI X

, ZHAO J

, GUO

, WANG H

, XING

. Cloning and functional analysis of the GmCTR1 in soybean
Scientia Agricultura Sinica, 2017,50(16):3082-3091. (in Chinese)

DOI:10.3864/j.issn.0578-1752.2017.16.003 URL [本文引用: 1]

茹京娜, 于太飞, 陈隽, 陈明, 周永斌, 马有志, 徐兆师, 闵东红 . 小麦锌指转录因子TaDi19A对低温的响应及其互作蛋白的筛选
中国农业科学, 2017,50(13):2411-2422.

DOI:10.3864/j.issn.0578-1752.2017.13.001 URL [本文引用: 1]

【目的】锌指类转录因子在植物逆境信号转导和非生物胁迫响应中发挥重要的作用。通过对小麦锌指转录因子基因TaDi19A的耐冷性能进行鉴定,利用酵母双杂交技术筛选并获得与TaDi19A互作的候选蛋白,以解析TaDi19A介导的抗逆调控机制。【方法】通过对低温处理的小麦转录组测序结果进行分析,获得一个锌指类转录因子TaDi19A。利用生物信息学的方法分析TaDi19A的分子特性,用SMART在线工具进行蛋白结构分析；用GSDS和PHYRE2在线工具分别对TaDi19A结构和蛋白三级结构进行分析；用NetPhos 2.0 Server数据库预测TaDi19A蛋白磷酸化位点。以低温处理的小麦cDNA作为模板,通过SYBR Green染料法进行实时荧光定量PCR,检测TaDi19A在低温处理不同时间段的表达模式。构建植物表达载体pBI121-TaDi19A,通过花序侵染法转化拟南芥,用T3代拟南芥进行耐冷性鉴定,分析低温处理对转基因拟南芥的根长、鲜重和存活率的影响。检测转TaDi19A拟南芥中抗逆相关基因表达变化,分析TaDi19A调控植物耐冷性的作用机制。构建诱饵载体pGBKT7-TaDi19A,验证自激活活性；利用酵母双杂交技术,将诱饵载体pGBKT7-TaDi19A和小麦cDNA文库共转化酵母AH109感受态细胞,通过SD/-Trp/-Leu/-His/-Ade和X-α-gal显蓝反应筛选得到阳性克隆,测序和BLAST分析获得候选蛋白。【结果】小麦TaDi19A编码区全长747 bp,编码248个氨基酸,分子量为28.03 kD,等电点为4.74,基因含4个外显子,3个内含子。TaDi19A蛋白靠近N-端包含锌指结合结构域,C端为Di19结构域,预测的TaDi19A蛋白三级结构包含2个α螺旋结构。磷酸化位点分析结果显示TaDi19A蛋白含有12个丝氨酸、9个苏氨酸和3个酪氨酸磷酸化位点。实时荧光定量PCR结果显示,TaDi19A受低温胁迫诱导表达。正常生长条件下,转基因和野生型拟南芥没有明显差异,低温处理下,转基因拟南芥的根长明显大于野生型拟南芥,并且耐冷性强于野生型拟南芥。下游基因检测结果表明,低温处理后,CBL1、CBL2和KIN1等冷胁迫响应相关基因在野生型和转基因植株中表达量都升高,在转基因植株中的表达量显著高于野生型,表明TaDi19A可能通过调节下游冷胁迫响应相关基因的表达提高转基因植物的耐冷性。通过对酵母双杂交系统筛选到的候选互作蛋白进行初步分析表明,这些候选互作蛋白主要参与植物体的信号转导和非生物胁迫响应过程,表明TaDi19A在植物的逆境信号转导及非生物胁迫响应过程中发挥着重要作用。【结论】小麦TaDi19A受低温诱导表达,过表达能够提高转基因拟南芥的耐冷性；而TaDi19A功能的发挥可能需要其他蛋白的参与。

RU J

, YU T

, CHEN

, ZHOU Y

, MA Y

, XU Z

, MIN D

. Response of wheat zinc-finger transcription factor TaDi19A to cold and its screening of interacting proteins
Scientia Agricultura Sinica, 2017,50(13):2411-2422. (in Chinese)

DOI:10.3864/j.issn.0578-1752.2017.13.001 URL [本文引用: 1]

侯思宇, 孙朝霞, 郭彬, 王玉国, 李贵全, 韩渊怀 . 大豆两个C2H2型转录因子基因序列特征及表达分析
植物生理学报, 2014,50(5):665-674.

[本文引用: 1]

HOU S

, SUN Z

, GUO

, WANG Y

, LI G

, HAN Y

. Cloning and expression analysis of two C2H2 transcription factors in soybean
Plant Physiology Journal, 2014,50(5):665-674. (in Chinese)

[本文引用: 1]

[40]

HUANG

, SUN S

, XU D

, YANG

, BAO Y

, WANG Z

, TANG H

, ZHANG H

. Increased tolerance of rice to cold, drought and oxidative stresses mediated by the overexpression of a gene that encodes the zinc finger protein ZFP245
Biochemical and Biophysical Research Communications, 2009,389(3):556-561.

DOI:10.1016/j.bbrc.2009.09.032 URL [本文引用: 1]

[41]

RAY P

, HUANG B

, TSUJI

. Reactive oxygen species (ROS) homeostasis and redox regulation in cellular signaling
Cell Signal, 2012,24(5):981-990.

DOI:10.1016/j.cellsig.2012.01.008 URL [本文引用: 1]

[42]

KORAMUTLA M

, KAUR

, NEGI

, VENKATACHALAM

, BHATTACHARYA

. Elicitation of jasmonate-mediated host defense in Brassica juncea (L.) attenuates population growth of mustard aphid Lipaphis erysimi (Kalt.)
Planta 2014,240(1):177-194.

DOI:10.1007/s00425-014-2073-7 URL [本文引用: 1]

The productivity of Brassica oilseeds is severely affected by its major pest: aphids. Unavailability of resistance source within the crossable germplasms has stalled the breeding efforts to derive aphid resistant cultivars. In this study, jasmonate-mediated host defense in Indian mustard Brassica juncea (L.) Czern. was evaluated and compared with regard to its elicitation in response to mustard aphid Lipaphis erysimi (Kalt.) and the defense elicitor methyl jasmonate (MeJ). Identification of jasmonate-induced unigenes in B. juncea revealed that most are orthologous to aphid-responsive genes, identified in taxonomically diverse plant-aphid interactions. The unigenes largely represented genes related to signal transduction, response to biotic and abiotic stimuli and homeostasis of reactive oxygen species (ROS), in addition to genes related to cellular and metabolic processes involved in cell organization, biogenesis, and development. Gene expression studies revealed induction of the key jasmonate biosynthetic genes (LOX, AOC, 12-OPDR), redox genes (CAT3 and GST6), and other downstream defense genes (PAL, ELI3, MYR, and TPI) by several folds, both in response to MeJ and plant-wounding. However, interestingly aphid infestation even after 24 h did not elicit any activation of these genes. In contrast, when the jasmonate-mediated host defense was elicited by exogenous application of MeJ the treated B. juncea plants showed a strong antibiosis effect on the infesting aphids and reduced the growth of aphid populations. The level of redox enzymes CAT, APX, and SOD, involved in ROS homeostasis in defense signaling, and several defense enzymes viz. POD, PPO, and PAL, remained high in treated plants. We conclude that in B. juncea, the jasmonate activated endogenous-defense, which is not effectively activated in response to mustard aphids, has the potential to reduce population growth of mustard aphids.

[43]

乔俊卿, 张心宁, 梁雪杰, 刘永锋, 刘邮洲 . 枯草芽孢杆菌PTS-394诱导番茄对灰霉病的系统抗性
中国生物防治学报, 2017,33(2):219-225.

[本文引用: 1]

QIAO J

, ZHANG X

, LIANG X

, LIU Y

. Plant system resistance triggered by root-colonizing Bacillus subtilis PTS-394 and its control effect on tomato gray mold
Chinese Journal of Biological Control, 2017,33(2):219-225. (in Chinese)

[本文引用: 1]

[44]

HAO Z

, WANG L

, HE Y

, LIANG J

, TAO R

. Expression of defense genes and activities of antioxidant enzyme in rice resistance to rice stripe virus and small brown planthopper
Plant Physiology and Biochemistry, 2011,49(7):744-751.

DOI:10.1016/j.plaphy.2011.01.014 URL [本文引用: 1]

The rice variety Tai06-1 is resistant to rice stripe disease and Xiushui63 is a highly susceptible rice variety to this disease. These two varieties were used to analyze the expression patterns of defense genes and antioxidant defense responses at the seedling stage, upon feeding with viruliferous small brown planthopper (SBPH) and nonviruliferous SBPH, respectively. The expression levels of CP (coat protein) gene of rice stripe virus (RSV) were higher upon feeding with viruliferous SBPH in Xiushui63 than in Tai06-1 throughout most of the experimental period, suggesting that RSV replicaiton is disturbed in Tai06-1 but not in Xiushui63, therefore, the resistance to RSV is higher in Tai06-1 than in Xiuhsui63. We found that defense genes PR1a (pathogenesis-related class la), PAL (phenylalanine ammonia-lyase), and CHS (chalcone synthase) may play roles in the defense responses to both RSV and SBPH in Tai06-1, and PR4 and PR10a may only participate in defending against SBPH attack but not against RSV infection in Tai06-1. Our data reveal that Gns1 (1,3; 1,4-13-glucanase) may participate in the defense responses to both RSV and SBPH in Xiushui63 but not in Tai06-1, and LOX (lipoxygenase) may only participate in defending against to SBPH in both Tai06-1 and Xiushui63. The antioxidant enzymes superoxide dismutase, peroxidase, catalase, hydrogen peroxide, and malondialdehyde coordinately participate in the resistance to RSV in Tai06-1, and that oxidative damage is less in Tai06-1 than in Xiushui63. (C) 2011 Elsevier Masson SAS.

[45]

孙璐, 黄瑞冬 . 高粱幼苗保护酶系统对盐胁迫的初期响应
沈阳农业大学学报, 2014,45(2):134-137.

[本文引用: 1]

SUN

, HUANG R

. Responses to salt stress of protective enzyme system in sorghum seedlings
Journal of Shenyang Agricultural University, 2014,45(2):134-137. (in Chinese)

[本文引用: 1]

[46]

李春燕, 徐雯, 刘立伟, 杨景, 朱新开, 郭文善 . 低温条件下拔节期小麦叶片内源激素含量和抗氧化酶活性的变化
应用生态学报, 2015,26(7):2015-2022.

URL [本文引用: 1]

以扬麦16和徐麦30为试验材料，利用人工气候室模拟低温逆境，研究拔节期-3 ℃和-5 ℃低温胁迫对小麦植株受冻率、叶片内源激素含量和抗氧化酶活性的影响.结果表明：随着处理温度的降低、胁迫时间的延长，小麦植株冻害等级与冻害指数增加，-5 ℃处理72 h两品种五级冻害率均为100%.低温处理结束当天，小麦叶片中内源激素脱落酸（ABA）、玉米素核苷（ZR）含量、抗氧化酶超氧化物歧化酶（SOD）、过氧化物酶（POD）、过氧化氢酶（CAT）活性随胁迫程度加重呈先升高后降低的趋势；处理结束后3 d，ABA、ZR含量及抗氧化酶活性较处理结束当天升高；至处理结束后6 d，与自然生长的对照处理接近.低温胁迫叶片中赤霉素（GA₃）含量下降，处理结束后3和6 d，扬麦16叶片中GA₃含量呈上升趋势，徐麦30则表现为先升高后下降.-5 ℃ 72 h重度胁迫处理叶片中ABA、ZR、GA₃含量和SOD、POD、CAT活性均较对照显著下降.相关分析表明，较高的ABA、ZR含量、SOD、POD、CAT活性以及较低的GA₃含量可减缓低温胁迫对小麦植株的伤害.

LI C

, XU

, LIU L

, YANG

, ZHU X

, GUO W

. Changes of endogenous hormone contents and antioxidative enzyme activities in wheat leaves under low temperature stress at jointing stage
Chinese Journal of Applied Ecology, 2015,26(7):2015-2022. (in Chinese)

URL [本文引用: 1]