作物杂志, 2023, 39(2): 83-90 doi: 10.16035/j.issn.1001-7283.2023.02.012

生理生化·植物营养·栽培耕作

水氮耦合对直播早籼稻苗期低温冷害的调控效应

王文霞,1, 吴自明2, 曾勇军2, 潘晓华2, 石庆华2, 曾研华,2

1吕梁学院生命科学系,033000,山西吕梁

2江西农业大学作物生理生态与遗传育种教育部重点实验室,330045,江西南昌

Regulatory Effects of Water-Nitrogen Coupling on Low Temperature Chilling Damage of Direct-Seeded Early Indica Rice at Seedling Stage

Wang Wenxia,1, Wu Ziming2, Zeng Yongjun2, Pan Xiaohua2, Shi Qinghua2, Zeng Yanhua,2

1Department of Life Sciences, Lüliang University, Lüliang 033000, Shanxi, China

2Key Laboratory of Crop Physiology Ecology and Genetic Breeding of Ministry of Education, Jiangxi Agricultural University, Nanchang 330045, Jiangxi, China

通讯作者: 曾研华,主要从事水稻高产栽培研究,E-mail:zyh74049501@163.com

收稿日期: 2022-11-12   修回日期: 2022-11-26   网络出版日期: 2023-02-20

基金资助: 国家自然科学基金(31760366)
国家重点研发计划(2017YFND0301605)
江西省杰出青年自然科学基金(20202ACBL215004)

Received: 2022-11-12   Revised: 2022-11-26   Online: 2023-02-20

作者简介 About authors

王文霞,主要从事作物栽培研究,E-mail:15797688640@163.com

摘要

为探究水氮耦合对直播早籼稻苗期低温冷害的调控作用,以耐冷品种湘早籼6号和冷敏感品种中嘉早17为供试材料,设置适温不施氮(CK)、低温不施氮(LT-0N)、低温低氮(LT-LN)、低温高氮(LT-HN)、低温淹水不施氮(LTF-0N)、低温淹水低氮(LTF-LN)和低温淹水高氮(LTF-HN)7个处理,处理5d,分析秧苗农艺性状、抗氧化酶活性、渗透调节物质、氮代谢相关酶与光合酶活性、内源激素含量等生理生态特性。结果表明,水氮耦合可缓解低温冷害对直播早籼稻造成的伤害。与LT-LN处理相比,LT-HN处理5d后冷敏感品种中嘉早17的苗高、根数、最大根长、鲜重、干重、顶部第1和第3片叶长显著增加;与LTF-0N处理相比,LTF-LN和LTF-HN处理5d后,不同耐性品种的根数、根长、鲜重和顶部第3片叶长显著增加,LTF-LN处理的根数和鲜重高于LTF-HN处理。与LT-0N处理相比,LT-HN和LTF-LN处理显著增加冷敏感品种中嘉早17叶片氮代谢相关酶和光合酶活性、硝态氮、叶绿素和内源生长促进类激素含量,但降低了叶片抗氧化酶活性、可溶性蛋白、渗透调节物质和内源生长抑制类激素含量,高氮和淹水低氮处理可对直播早籼稻苗期低温冷害起到缓解作用。

关键词: 直播早籼稻; 低温; 苗期; 水氮耦合; 生理特性

Abstract

In order to study the control methods of water-nitrogen coupling on low temperature chilling damage of direct-seeded early indica rice at the seedling stage, the cold-tolerant cultivar Xiangzaoxian 6 and cold- sensitive cultivar Zhongjiazao 17 were used as materials, and seven treatments including ambient temperature no nitrogen (CK), low temperature plus no nitrogen (LT-0N), low temperature plus low nitrogen (LT-LN), low temperature plus high nitrogen (LT-HN), low temperature flooding plus no nitrogen (LTF-0N), low temperature flooding plus low nitrogen (LTF-LN) and low temperature flooding plus high nitrogen (LTF-HN) were established for five days to analyse the physiological characteristics such as agronomic characters, antioxidant enzyme activities, osmotic regulation substances, nitrogen metabolism-related enzymes and photosynthase activities, endogenous hormone contents of rice seedlings. The results showed that water-nitrogen coupling treatments could alleviate the injury caused by low temperature chilling damage to direct-seeded early indica rice, compared with LT-LN treatment, the seedling height, root number, maximum root length, fresh weight, dry weight, T1 (the first leaf from top) leaf length and T3 (the 3rd leaf from top) leaf length of cold-sensitive cultivar Zhongjiazao 17 significantly increased after five days of LT-HN treatment; compared with LTF-0N treatment, the root number, root length, fresh weight and T3 leaf length of different tolerant cultivars increased significantly after five days of LTF-LN and LTF-HN treatments, the number of roots and fresh weight were higher in LTF-LN than that in LTF-HN. Compared with LT-0N treatment, LT-HN and LTF-LN treatments significantly increased the nitrogen metabolism-related enzymes, photosynthase activities, nitrate nitrogenin, chlorophyll and endogenous growth-promoting hormone contents in leaves of the cold-sensitive cultivar Zhongjiazao 17, but decreased leaf antioxidant enzyme activities, soluble protein, malonic dialdehyde, and endogenous growth- inhibiting hormones contents, high nitrogen and flooding low nitrogen treatments could alleviate low temperature cold damage in seedlings of direct-seeded early indica rice.

Keywords: Direct-seeded early indica rice; Low temperature; Seedling stage; Water-nitrogen coupling; Physiological traits

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本文引用格式

王文霞, 吴自明, 曾勇军, 潘晓华, 石庆华, 曾研华. 水氮耦合对直播早籼稻苗期低温冷害的调控效应. 作物杂志, 2023, 39(2): 83-90 doi:10.16035/j.issn.1001-7283.2023.02.012

Wang Wenxia, Wu Ziming, Zeng Yongjun, Pan Xiaohua, Shi Qinghua, Zeng Yanhua. Regulatory Effects of Water-Nitrogen Coupling on Low Temperature Chilling Damage of Direct-Seeded Early Indica Rice at Seedling Stage. Crops, 2023, 39(2): 83-90 doi:10.16035/j.issn.1001-7283.2023.02.012

水稻是世界上近30亿人口的主要粮食,提高水稻产量是保障粮食安全的有效途径[1]。我国每年因低温冷害平均损失稻谷30亿~500亿kg[2]。长江中下游、华南和西北水稻种植区经常会面临不同程度的低温胁迫,影响直播早稻高产稳产。江西省雨量充足,春季阴冷多雨,早稻苗期低温冷害及阴雨天气普遍存在,直播稻苗期遭遇“倒春寒”气候及低温冷害后出现大面积的烂种烂秧,导致秧苗生长迟缓,产量降低[3]。因此,减少烂种烂秧对直播稻的抗逆稳产栽培具有重要意义。生产中一般通过以水调温、增施氮肥等措施降低低温对直播稻的危害或直接提高直播稻耐冷性。农民经常试图通过浅层淹水的措施来减少低温对早稻秧苗的损害,从而提高秧苗成活率,降低直播稻的生产风险。此外,还可通过增施氮肥缓解低温冷害对直播稻生长发育及产量的影响[4]。但是,这种实际的生产措施缺乏理论依据支撑。氮肥的投入是直播稻增产的重要手段之一,为了获得高产,部分稻田年平均施氮量已高达500kg/hm2以上[5],过量的氮肥不仅增加了生产成本,而且加剧了环境污染[6]。如何通过淹水及氮素高效利用来缓解低温对直播稻生产的影响、降低高氮对环境造成的污染,已成为直播稻生产中亟待解决的问题。周永进[4]通过设置提早和正常直播发现,在2年大田直播稻生产中,高氮处理在提前播种下的产量比低氮处理分别增加了11.8%和11.7%。向镜等[7]研究表明,苗期低温淹水条件下株高显著增加,胚芽鞘增长加快,可有效降低死苗率。说明施氮或淹水有利于减少低温对水稻造成的伤害。以往的研究主要以淹水或氮肥施用量单因素缓解低温冷害为主,而关于低温、淹水及氮肥对直播早籼稻苗期叶片生理特性的复合缓解效应研究报道较少。本研究以耐冷品种湘早籼6号和冷敏感品种中嘉早17为材料,于低温处理下设置不同氮肥施用量和淹水处理,系统研究低温冷害下水氮耦合对直播早籼稻幼苗生长及生理生态特性的影响,明确直播早籼稻苗期低温冷害下水、氮措施的调控作用,为南方稻区直播稻生产与抗逆栽培提供理论依据。

1 材料与方法

1.1 试验材料

在前期试验[3]基础上,供试材料选用冷敏感品种中嘉早17(由江西现代种业股份有限公司提供)和耐冷品种湘早籼6号(由湖南省农业科学院提供)。盆栽用土取自江西农业大学科技园水稻试验田表层(0~20cm)土壤,试验用盆高15.0cm、长25.0cm、宽23.0cm。试验土壤pH 5.52、有机质33.95g/kg、全氮2.03g/kg、速效磷24.78mg/kg、速效钾99.42mg/kg。土壤自然风干,用土壤研磨机粉碎,然后过100目筛。每盆装6kg风干土,直播前在水中浸泡2周,直播前1d施复合肥(N-P-K= 15-15-15)3g。

1.2 试验设计

盆栽试验于2019年在江西农业大学作物生理生态与遗传育种教育部重点实验室人工气候室(气候室上下2层,共32m2)进行,设置适温不施氮(25°C,CK)、低温不施氮(8°C,LT-0N)、低温低氮(8°C,120kg/hm2,折合1.57g/盆,LT-LN)、低温高氮(8°C,180kg/hm2,2.5g/盆,LT-HN)、低温淹水不施氮(8°C,5~6cm水层,LFT-0N)、低温淹水低氮(8°C,5~6cm水层,120kg/hm2,1.57g/盆,LFT-LN)、低温淹水高氮(8°C,5~6cm水层,180kg/hm2,2.5g/盆,LFT-HN)共7个处理。各处理光照均为12h(7:00-19:00),光照强度为100mol/(m∙s),人工气候室相对湿度为75%。选取刚破胸露白的种子进行播种,每盆手动点播40粒种子,直播后置于25°C(27°C/23°C)人工气候室中,待其生长到2叶1心后,选取出苗率达95%的盆栽进行不同处理,于低温处理前施氮肥,施肥后第2天移动到8°C(10°C/6°C)人工气候室进行低温处理5d。在处理过程中,每个盆栽的位置不定时地旋转,以避免光照不均对幼苗生长的影响。3次重复,每次重复设20盆。于水氮耦合及低温处理5d后对水稻倒2叶进行取样,每个处理取样3次重复。除用于农艺性状调查的样本外,所有样品均在液氮中快速冷冻,保存于-80°C超低温冰箱,以待分析样品的生理特性。

1.3 测定项目与方法

1.3.1 农艺性状

水氮耦合处理5d后取长势均匀的秧苗30株,每个处理3次重复,将秧苗洗净擦干后测量鲜重、苗高、顶部第1、第2和第3片叶长,记录每株秧苗的根数和最大根长,然后将秧苗分别装于网袋,置于烘箱中80°C烘干至恒重后测量干重。

1.3.2 抗氧化酶活性及可溶性蛋白含量

采用李合生[8]的方法测定超氧化物歧化酶(SOD)、过氧化物酶(POD)及过氧化氢酶(CAT)活性。称取0.1g叶片,加入1mL提取液(磷酸缓冲液),进行冰浴匀浆,在8000转/min、4°C下离心10min后取上清液置于冰上待测。用考马斯亮蓝比色法[8]测定可溶性蛋白含量。

1.3.3 叶片膜脂过氧化物

采用硫代巴比妥酸法测定丙二醛(MDA)含量。称取0.1g叶片,加入1mL提取液(磷酸缓冲液),进行冰浴匀浆,在8000转/min、4°C下离心10min,取上清液置于冰上待测。采用茚三酮比色法测定游离脯氨酸(Pro)含量[9],称取0.1g叶片,加入1mL提取液(磺基水杨酸溶液),进行冰浴匀浆,之后置于90°C振荡提取10min,常温10 000转/min离心10min后,取上清液,置冰上待测。

1.3.4 氮代谢相关酶

采用苏州科铭生物技术有限公司生产的试剂盒测定硝酸还原酶(NR)、谷氨酰胺合成酶(GS)、谷氨酸脱氢酶(GDH)活性和硝态氮含量[10-11]

1.3.5 光合酶活性

用80%的丙酮溶液提取叶绿素,采用分光光度法测定叶绿素含量[8]。采用科铭生物有限公司生产的试剂盒测定核酮糖-1,5-二磷酸羧化酶(Rubisco)活性[10-11]

1.3.6 内源激素含量

采用高效液相色谱法(HPLC)测定脱落酸(ABA)和赤霉素(GA3)等内源激素含量[12]。测定方法略作调整:称取0.1g叶片,放入研钵中磨碎,加入1mL试剂I(80mL甲醇+20mL水+1mL乙酸),4℃下浸提过夜。8000转/min下离心10min,离心后取出上清液,氮吹至水相。用试剂II(柠檬酸)调pH至2~3,用乙酸乙酯萃取3次,合并乙酸乙酯相,氮吹至干。甲醇定容至0.5mL,针头式过滤器过滤于带有内衬管的样品瓶内待测。

1.4 数据处理

采用Excel 2010进行数据分析和整理,使用SAS进行方差分析,用HSD检验法进行差异显著性检验(P<0.05)。

2 结果与分析

2.1 水氮耦合对直播早籼稻苗期低温冷害农艺性状的影响

表1可知,品种和水氮耦合处理对根数、最大根长、鲜重、顶部第1和第3片叶长均有极显著影响,品种与水氮耦合处理的互作效应对苗高、最大根长、鲜重、干重、顶部第2和第3片叶长有显著或极显著影响。低温冷害胁迫下,与LT-0N处理相比,LT-LN和LT-HN处理5d后不同耐性品种的根数、最大根长、鲜重、顶部第2和第3片叶长显著增加,高氮处理的增幅大于低氮处理,高氮处理5d后冷敏感品种中嘉早17的苗高、根数、最大根长、鲜重、干重、顶部第1和第3片叶长较低氮处理显著增加。此外,通过淹水处理,LTF-LN和LTF- HN处理与LTF-0N处理相比,不同耐性品种的根数、根长、鲜重和顶部第3片叶长显著增加,苗高、干重、顶部第1和第2片叶长也呈增加趋势,但低氮处理的根数和鲜重高于高氮处理,苗高无差异。

表1   水氮耦合对直播早籼稻苗期低温冷害农艺性状的影响

Table 1  Effects of water-nitrogen coupling on agronomic characteristic of direct-seeded early indica rice under low temperature chilling damage condition at the seedling stage

品种
Variety
处理
Treatment
苗高
Seedling
height (cm)
根数
Root
number
最大根长
Maximum root
length (cm)
鲜重
Fresh weight
(mg/100 plants)
干重
Dry weight
(mg/100 plants)
叶长Leaf length (cm)
T1T2T3
湘早籼6号
Xiangzaoxian 6
CK35.2a12.7b3.2c283.3a48.3b2.5c10.2bc21.3bc
LT-0N27.4d9.7d1.8e195.7f38.8d2.3d9.3d19.5d
LT-LN28.5cd11.3c2.3d235.7de44.3c2.5bc10.7ab20.8c
LT-HN30.2bc13.3ab2.4d257.7bc48.3b2.6bc11.0ab21.6bc
LTF-0N28.3cd10.0d2.4d220.3e42.7c2.6bc9.7cd20.8c
LTF-LN32.1b14.3a4.1a274.7ab50.5ab2.8b11.1a23.0a
LTF-HN28.9cd12.3bc3.9b252.0cd52.9a3.1a9.4d22.2ab
中嘉早17
Zhongjiazao 17
CK31.6a12.0c3.2ab275.0ab43.2bc2.5bc9.9bc19.5bc
LT-0N25.7d9.7d1.6d231.0d40.7c2.0d8.9d17.1e
LT-LN27.8c13.0bc2.2c259.3bc42.5bc2.2d10.4bc19.1cd
LT-HN30.3ab15.0a2.9b288.3a50.9a2.4bc11.0ab20.3b
LTF-0N28.9bc11.7c2.3c241.3cd47.7ab2.4c9.4cd18.3d
LTF-LN31.1a15.3a3.2ab277.7ab51.3a2.6ab10.4bc19.5bc
LTF-HN31.3a14.0ab3.4a266.7ab52.1a2.7a11.5a21.5a
V2.515.5**13.7**25.9**0.224.7**0.0140.8**
T24.3**28.8**112.5**31.5**26.6**19.3**11.1**27.5**
V×T4.1**2.08.3**3.0*3.4*1.35.8**4.0**

T1:顶部第1片叶;T2:顶部第2片叶;T3:顶部第3片叶;V:品种;T:处理。同列同一品种数据后不同小写字母表示差异达显著水平(P < 0.05)。“*”和“**”分别表示5%和1%水平影响显著

T1: the 1st leaf from top; T2: the 2nd leaf from top; T3: the 3rd leaf from top; V: variety; T: treatment. Different lowercase letters after the data of the same cultivar in the same column indicate a significant level of difference (P < 0.05).“*”and“**”indicate significant effects at 0.05 and 0.01 levels, respectively

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2.2 水氮耦合对直播早籼稻苗期低温冷害抗氧化酶活性及可溶性蛋白含量的影响

图1可知,与CK处理相比,LT-0N、LT-LN和LT-HN处理5d后,不同耐性品种的抗氧化酶(SOD和CAT)活性和可溶性蛋白含量显著增加。低温冷害胁迫下,与不施氮相比,低氮和高氮处理5d后SOD、CAT活性和可溶性蛋白含量呈降低趋势,高氮处理较低氮处理显著降低不同耐性品种的CAT活性和可溶性蛋白含量。通过淹水处理,LTF-LN处理较LTF-0N处理显著降低了不同耐性品种的SOD活性和可溶性蛋白含量,LTF-HN处理呈增加趋势。

图1

图1   水氮耦合对直播早籼稻苗期低温冷害抗氧化酶活性和可溶性蛋白含量的影响

不同小写字母表示在P < 0.05水平上差异显著,同一品种进行统计分析,下同

Fig.1   Effects of water-nitrogen coupling on antioxidant enzyme activity and soluble protein content of direct-seeded early indica rice under low-temperature chilling damage condition at the seedling stage

Various lowercase letters above the bars mean significant difference at 0.05 probability level, statistical analysis of the same cultivar, the same below


2.3 水氮耦合对直播早籼稻苗期低温冷害叶片膜脂过氧化物的影响

图2可知,与CK处理相比,LT-0N、LT-LN和LT-HN处理5d后不同耐性品种的MDA和Pro含量显著增加,冷敏感品种中嘉早17的增加幅度大于耐冷品种湘早籼6号。低温冷害胁迫下,低氮和高氮处理较低温不施氮处理显著降低了不同耐性品种的MDA含量,低氮处理平均降幅为18.89%,高氮处理平均降幅为31.82%,高氮处理的降幅大于低氮处理,而Pro含量显著增加。此外,淹水处理后,LTF-LN处理较LTF-0N处理显著降低冷敏感品种中嘉早17的MDA含量,接近于CK处理,而Pro含量增加。

图2

图2   水氮耦合对直播早籼稻苗期低温冷害膜脂过氧化物的影响

Fig.2   Effects of water-nitrogen coupling on membrane peroxides of direct-seeded early indica rice under low-temperature chilling damage condition at the seedling stage


2.4 水氮耦合对直播早籼稻苗期低温冷害氮代谢相关酶活性的影响

图3可知,与CK处理相比,LT-0N、LT-LN和LT-HN处理5d后不同耐性品种的氮代谢相关酶(NR、GS、GDH)活性显著增加。低温冷害胁迫下,低氮和高氮处理较不施氮处理显著增加冷敏感品种中嘉早17的NR和GS活性,其中高氮处理的增幅大于低氮处理。此外,通过淹水处理,LTF-HN处理较LTF-0N处理显著增加不同耐性品种的NR、GS、GDH活性和硝态氮含量。

图3

图3   水氮耦合对直播早籼稻苗期低温冷害氮代谢相关酶活性的影响

Fig.3   Effects of water-nitrogen on the activities of nitrogen metabolism-related enzymes activity of direct-seeded early indica rice under low-temperature chilling damage condition at the seedling stage


2.5 水氮耦合对直播早籼稻苗期低温冷害光合酶活性的影响

图4可知,与CK处理相比,LT-0N处理5d后不同耐性品种的Rubisco活性和叶绿素含量显著降低,Rubisco活性降幅分别为20.63%和15.98%,叶绿素含量降幅分别为45.29%和43.89%。低温冷害胁迫下,LT-HN处理较LT-0N处理显著增加不同耐性品种的Rubisco活性和叶绿素含量;此外,通过淹水处理,与LTF-0N处理相比,LTF-LN和LTF-HN处理也显著增加Rubisco活性和叶绿素含量,其中高氮处理的增幅效应大于低氮处理。

图4

图4   水氮耦合对直播早籼稻苗期低温冷害光合酶活性的影响

Fig.4   Effects of water-nitrogen on photosynthetic enzyme activities of direct-seeded early indica rice under low-temperature chilling damage condition at the seedling stage


2.6 水氮耦合对直播早籼稻苗期低温冷害下内源激素含量的影响

图5可知,与CK处理相比,LT-0N和LT-LN处理5d后不同耐性品种的ABA含量显著增加,而IAA含量显著降低。低温冷害胁迫下,LT-HN处理较不施氮处理显著降低冷敏感品种中嘉早17的ABA含量,降幅为32.26%,而不同耐性品种的IAA含量显著增加。此外,淹水处理后,LTF-LN处理较LTF-0N处理ABA含量显著降低,IAA含量显著增加。

图5

图5   水氮耦合对直播早籼稻苗期低温冷害内源激素含量的影响

Fig.5   Effects of water-nitrogen coupling on endogenous hormone content of direct-seeded early indica rice under low-temperature chilling damage condition at the seedling stage


3 讨论

3.1 水氮耦合对直播早籼稻苗期低温冷害农艺性状的缓解效应

温度、水分和氮素是影响水稻生长发育的重要因素。低温使得直播稻出现大面积的烂种烂芽,导致幼苗出现黄叶、卷叶和死苗现象,严重影响直播稻生产[13-14]。前人[15-16]研究表明,在低温冷害期,灌深水可起到增温保温作用,低温过后及时排水露田,可达到防寒效果。邹江石等[17]研究得出,15~ 20cm灌溉层可缓解低温冷害的伤害,通过增加水稻冠层温度,起到“以水增温”的效果。氮肥施用可促进幼苗生长,具有明显的增产效果,是植物生长中重要的肥料[18]。但与此同时,过量增施氮肥也会增加土壤氮素残留量,过量的氮肥流失到土壤环境中去,使地下水体遭受污染,江河湖泊出现富营养化[19]。因此,合理的氮肥用量既有利于水稻产量增加又可以保护生态环境。本研究结果表明,低温冷害胁迫5d后,低氮和高氮处理可显著增加不同耐性品种的苗高、根数、根长、鲜重和叶长,以高氮处理增幅明显,施氮可缓解低温对直播早稻秧苗农艺性状的不利影响。这与前人[20]关于低温及施氮处理对水稻农艺性状研究结果相似。低温施氮可促进水稻生长恢复,刺激了细胞的分裂和伸长,随施氮量增加,水稻氮吸收量和干物质积累均不同程度增加。此外,通过淹水低氮和淹水高氮处理,秧苗根数、根长和鲜重显著增加,苗高、干重和叶长也呈增加趋势。浅层淹水使植物体内乙烯含量提高,促进了细胞分裂和伸长,从而促进水稻株高的增大[21],低温淹水低氮处理下直播早籼稻表现出较好的适应性,可缓解低温胁迫对直播早籼稻秧苗性状的影响,说明低温胁迫下,水氮耦合既可以减少氮肥的施用量,又可以有效缓解低温对直播早籼稻幼苗农艺性状造成的伤害。

3.2 水氮耦合对直播早籼稻苗期低温冷害抗氧化酶系统的保护作用

当农业生产遭遇逆境胁迫时,水分灌溉和氮肥的施用有利于通过调控植株生理特性、叶片酶活性和代谢水平提高水稻的耐受力[22]。研究[23]表明,淹水处理后施用氮肥,水稻生长得以恢复,这是因为淹水胁迫处理结束后植株恢复到自然环境中生长,活性氧含量虽然升高,但是在抗氧化酶的作用下又可以逐渐恢复,淹水后施氮调节了植物的生理生化过程,使植株能够快速恢复。本文在前人研究的基础上开展了关于低温胁迫下水氮耦合对直播早籼稻幼苗的影响,结果发现,低温冷害下,高氮处理显著降低CAT活性和可溶性蛋白含量,说明在施用氮肥后直播早籼稻植株体内生理代谢水平逐渐恢复,达到调控生理生化过程从而提高其抗性和恢复生长的功能。同时,通过淹水低氮和淹水高氮处理,使得抗氧化酶(SOD、POD、CAT)活性和可溶性蛋白含量呈先降低后增加的趋势,这可能是由于过多的施氮量反而破坏了细胞内氮代谢平衡,导致活性氧产生。

此外,当直播早籼稻幼苗遭遇低温冷害时,可通过渗透调节过程来维持细胞内外的渗透平衡,这样就会导致参加渗透调节反应的物质发生变化,避免植株受冷害损伤,起到提高植物耐低温胁迫能力的作用[24]。MDA含量可反映植物细胞膜脂过氧化水平,以及低温对细胞膜结构伤害程度的大小[25]。本研究表明,低温冷害下,低氮和高氮处理显著降低了不同耐性品种的MDA含量,表明施氮有助于缓解膜脂过氧化酶在膜脂过氧化中的参与,降低低温冷害胁迫对细胞膜系统的损伤,减少细胞内渗透压力。同时,通过淹水低氮处理亦可显著降低冷敏感品种中嘉早17的MDA含量,接近于对照,而Pro含量增加。进一步说明淹水配施低氮更有利于减少低温胁迫对直播早籼稻叶片生理活性的影响,保持叶片结构的完整性。

3.3 水氮耦合对直播早籼稻苗期低温冷害氮代谢相关酶活性的调节效应

NR是水稻进行氮代谢的起始酶,也是硝酸还原过程的调节限速酶,该酶活性受到生长生育进程的影响,而且对温度、水分和氮素极其敏感[26]。植株抗逆性生理研究发现,NR活性的大小与其抵抗逆境胁迫的能力有关。GS是氮代谢途径中另外一种限速酶,可以使95%的无机氮转化为有机氮,达到既满足对氮元素的需求,又可以使植株免受氨中毒影响[27]。陶乐圆[28]研究表明,高氮处理时氮代谢相关酶活性增加较为显著,高氮使植株有更好的氮代谢能力。本研究得出,低温条件下,低氮和高氮处理显著增加冷敏感品种中嘉早17的NR和GS活性,这可能与细胞膜透性增加有关,NR活性的增加使植物体内含氮化合物的含量提高,可用来抵抗低温逆境胁迫。该研究结果与陶乐圆[28]的研究结果较为相似。而高氮的增幅效应大于低氮处理,这说明在低氮处理下,植物叶片的氮同化力较弱,碳代谢相对较强,而高氮处理下,硝酸还原能力增强,叶片光合产物逐渐转向氮代谢,使氮代谢能力增强。此外,本研究结果表明,低温冷害下,淹水高氮处理可显著增加不同耐性品种的NR、GS、GDH活性和硝态氮含量。说明适宜的施氮量和淹水管理措施提高了直播早籼稻幼苗应对低温胁迫的适应性,可减缓低温对幼苗的损伤。

3.4 水氮耦合对直播早籼稻苗期低温冷害光合作用和内源激素含量的调控作用

水稻的光合作用受温度和水分的影响很大。在营养生长期,随着温度的降低,植株的活力逐渐减弱,水分与养分吸收能力呈降低趋势,光合作用能力减弱[29]。Rubisco活性的大小直接影响植株光合作用能力的强弱。Makino等[30]研究认为,低温下随着叶片含氮量的提高,光合作用相关酶活性提高,植株的光合能力增强。本研究结果也得出,经低温高氮处理5d后,Rubisco活性和叶绿素含量较低温不施氮处理显著增加,说明高氮在一定程度上可缓解低温对直播早籼稻光合作用的抑制。胡凝[31]研究表明,当遭遇阴雨天气时,采用深水护苗可降低低温冷害对水稻植株的伤害,减少对产量造成的损失。本研究结果表明,低温处理下,淹水高氮处理显著增加Rubisco活性和叶绿素含量,说明在一定程度上减缓了低温胁迫对直播早籼稻幼苗叶片光合酶活性的伤害,改善叶片功能。而耐冷品种湘早籼6号自身反应机制有利于抵抗低温冷害对叶片光合作用造成的伤害。

水氮耦合对直播早籼稻苗期低温冷害的调控效应涉及ABA、IAA等多种内源激素的综合调控[32-33]。遭遇低温时,细胞中IAA含量降低,抑制植株生长,促进气孔关闭,降低蒸腾作用以适应环境。本研究表明,低温胁迫下,高氮处理使冷敏感品种中嘉早17的内源生长抑制类激素(ABA)含量显著降低,但内源生长促进类激素(IAA)含量增加。这与余小芬等[34]的研究结果相似。说明施氮对低温胁迫下直播早籼稻幼苗叶片内源激素具有较好的调节恢复效应。同时,本研究还表明,淹水低氮处理也显著降低了ABA含量,且显著增加了IAA含量。表明高氮和淹水低氮作用可对直播早籼稻幼苗低温冷害起到缓解作用。

4 结论

与低温低氮处理相比,低温高氮处理5d显著增加冷敏感品种中嘉早17的苗高、根数、最大根长、鲜重、干重、顶部第1和第3片叶长;同时,与低温淹水不施氮相比,低温淹水低氮和高氮处理5d后,不同耐性品种的根数、根长、鲜重和顶部第3片叶长显著增加,低温淹水低氮处理的根数和鲜重高于低温淹水高氮处理,苗高无差异。此外,与低温不施氮处理相比,低温高氮和低温淹水低氮处理增加了叶片氮代谢相关酶、光合酶活性和IAA含量,但降低了叶片抗氧化酶活性、可溶性蛋白、渗透调节物质和内源生长抑制类激素(ABA)含量,低温高氮和低温淹水低氮可对直播早籼稻苗期低温冷害起到缓解作用。总体来说,低温条件下,采用高氮处理可缓解低温冷害对直播早籼稻秧苗造成的伤害;而采用淹水灌溉低氮处理既可以减少氮肥施用量,又能对直播早籼稻苗期低温冷害起到缓解作用。因此,淹水低氮处理对直播早籼稻苗期低温冷害的缓解效果最好。

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中国水稻科学, 2016, 30(5):525-531.

DOI:10.16819/j.1001-7216.2016.5183      [本文引用: 1]

为了研究淹涝水温对具有不同耐淹能力水稻材料形态和生理的影响,明确淹涝水温对水稻幼苗耐涝能力影响的机理,采用盆栽试验,选取导入耐淹基因的耐淹品种IR64Sub1和淹水敏感品种IR42以及耐淹能力较强的品种冈优725和泰优398,设计20℃和30℃两种水温进行淹水处理,淹水时间分为0, 3, 6, 9和12 d。结果表明较低淹水温度下,水中的溶氧量显著高于高水温处理;随着淹水时间延长,高水温处理溶氧量下降速度显著快于低水温处理。高水温条件下水稻幼苗株高增长率要显著高于低水温处理,虽然高水温处理下耐涝品种株高增长率也显著增加,但是增长量要显著低于其他材料。高水温处理水稻茎鞘内可溶性糖和淀粉消耗高于低水温处理,但随着淹水时间的延长两种温度处理间差异逐渐变小,较低的水温可以减少叶片叶绿素降解速率。因此,降低淹水温度可以提高淹水条件下水稻幼苗的成活率。以上结果揭示了低水温可以减少茎鞘的伸长,减少非结构性碳水化合物的消耗及叶绿素的降解,从而水稻淹涝后能维持较高的成活率。这些发现有助于理解较低洪涝水温下水稻存活更长时间的机理。

李合生. 植物生理生化实验原理和技术. 北京: 高等教育出版社, 2006.

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张志良. 植物生理学实验指导. 北京: 高等教育出版社,1990.

[本文引用: 1]

Chen G, Zhang Y, Ruan B P, et al.

OsHAK1 controls the vegetative growth and panicle fertility of rice by its effect on potassium-mediated sugar metabolism

Plant Science, 2018, 274:261-270.

DOI:S0168-9452(18)30352-2      PMID:30080612      [本文引用: 2]

Plant growth and reproduction are both energy-requiring processes; the necessary energy is supplied by the products of photosynthesis. Both the vegetative growth and reproductive success of rice are compromised by the absence of a functional copy of the gene OsHAK1. Here, a comparison between wild type rice and OsHAK1 knockout mutants not only confirmed the known detrimental effect of the absence of OsHAK1 on root growth, pollen viability and fertility, but also showed that sucrose phosphate synthase activity was lowered, and the sucrose content of the leaves was markedly increased, due to a partial block on the up-loading of sucrose into the phloem. The impaired allocation of sugar to the roots and spikelets caused by the knocking out of OsHAK1 was accompanied by a down-regulation in the leaf sheaths and panicle axes of genes encoding sucrose transporters (SUT genes), which are active in the phloem, as well as in the roots and spikelets of those encoding monosaccharide transporters (MST genes), which transport hexose sugars across the plant plasma membrane. The activity of sucrose synthase, acid invertase and neutral invertase in the roots of mutant plants assayed at the tillering stage, and in their spikelets, assayed during grain-filling, was significantly lower than in the equivalent organs of wild type plants. As a result, the supply of total soluble sugar, glucose and fructose to sink organs was reduced, consistent with the effect of the mutation on root growth and panicle fertility. Compared to wild type plants, the mutants accumulated less potassium (K) throughout the plant. The conclusion was that the failure to fully supply the demand of the mutant's sink organs for assimilate was responsible for its compromised phenotype, and that the deficiency in K uptake induced by the loss of OsHAK1 functionality was responsible for the disruption of sugar metabolism.Copyright © 2018 Elsevier B.V. All rights reserved.

Wu K, Li J X, Luo J P, et al.

Effects of elevated CO2 and endophytic bacterium on photosynthetic characteristics and cadmium accumulation in Sedum alfredii

Science of the Total Environment, 2018, 643:357-366.

DOI:10.1016/j.scitotenv.2018.06.131      URL     [本文引用: 2]

Teng Y B, Cui H Q, Wang M Y, et al.

Nitrate reductase is regulated by circadian clock-associated 1 in Arabidopsis thaliana

Plant and Soil, 2017, 416(1):1-9.

DOI:10.1007/s11104-017-3183-3      URL     [本文引用: 1]

王慰亲. 种子引发促进直播早稻低温胁迫下萌发出苗的机理研究. 武汉:华中农业大学, 2019.

[本文引用: 1]

王亚男, 范思静.

低温胁迫对水稻幼苗叶片生理生化特性的影响

安徽农业科学, 2017, 45(5):8-9.

[本文引用: 1]

雷俊, 冯敏玉.

2011年南昌市影响粮食生产的气象灾害

天津农业科学, 2012, 18(6):131-134.

[本文引用: 1]

彭春瑞, 刘小林, 李名迪, .

江西水稻主要气象灾害及防御对策

江西农业学报, 2005, 17(4):127-130.

[本文引用: 1]

邹江石, 吕川根, 姚克敏, .

两系法杂交稻安全制种的低温防御灌水理论与技术

中国农业科学, 2005, 39(9):1780-1786.

[本文引用: 1]

曹超豪, 沈天花, 施翔, .

不同氮水平下晚稻品种响应抽穗扬花期低温的产量及生理差异

华北农学报, 2020, 35(2):178-186.

DOI:10.7668/hbnxb.20190343      [本文引用: 1]

为探究不同氮水平下晚稻抽穗扬花期低温的产量及生理差异,以3个晚稻品种黄华占、荣优225、H优518为材料,设置低氮,N1;正常氮,N2;高氮,N3;超高氮,N4 4个处理。结果表明,低温条件下,3个品种不同氮肥处理的单株产量以N2处理最高,且H优518的单株产量最高,其次为荣优225,黄华占最低;3个品种在N1处理下的产量均最低,主要是单株有效穗较低导致的;不同品种N3、N4的单株产量低于N2是由于结实率较低所导致。3个品种穗部干物质积累N3处理较N2处理均有不同幅度上升,其中以H优518上升幅度最小。低温后,N2处理的包颈率以黄华占最高。低温来临后,3个品种叶片叶绿素含量及净光合速率的下降幅度随着施氮量的增加而增大,蒸腾速率、气孔导度也表现出类似特点。正常氮处理下,H优518叶片丙二醛(MDA)含量和过氧化物酶(POD)活性的上升幅度最小,游离脯氨酸含量上升幅度最大,但过氧化氢酶(CAT)和超氧化物歧化酶(SOD)活性及可溶性蛋白含量均不同程度下降,H优518降幅最小。晚稻在抽穗扬花期遇低温时产量会降低,少施氮肥虽可使水稻株产生一定的抗性效应,但产量依然最低,高氮或超高氮会使植株抵抗力严重下降,导致产量降低,且不同品种存在一定的差异性。

宋勇生, 范晓晖, 林德喜, .

太湖地区稻田氨挥发及影响因素的研究

土壤学报, 2004, 41(2):265-269.

[本文引用: 1]

彭显龙, 刘元英, 罗盛国, .

实地氮肥管理对寒地水稻干物质积累和产量的影响

中国农业科学, 2006, 39(11):2286-2293.

[本文引用: 1]

杨生龙, 贾志英.

不同水分处理对水稻和旱稻产量及产量构成因子的影响

安徽农业科学, 2010, 38(31):17410-17412.

[本文引用: 1]

Gautam P, Lal B, Tripathi R, et al.

Impact of seedling age and nitrogen application on submergence tolerance of Sub1 and non-Sub 1 cultivars of rice (Oryza sativa L.)

Journal of Plant Growth Regulation, 2017, 36(3):629-642.

DOI:10.1007/s00344-016-9661-7      URL     [本文引用: 1]

Fukao T, Xu K, Ronald P C, et al.

A variable cluster of ethylene response factor-like genes regulates metabolic and developmental acclimation responses to submergence in rice

The Plant Cell, 2006, 18(8):2021-2034.

DOI:10.1105/tpc.106.043000      URL     [本文引用: 1]

Submergence-1 (Sub1), a major quantitative trait locus affecting tolerance to complete submergence in lowland rice (Oryza sativa), contains two or three ethylene response factor (ERF)–like genes whose transcripts are regulated by submergence. In the submergence-intolerant japonica cultivar M202, this locus encodes two ERF genes, Sub1B and Sub1C. In the tolerant near-isogenic line containing the Sub1 locus from the indica FR13A, M202(Sub1), the locus additionally encodes the ERF gene Sub1A. During submergence, the tolerant M202(Sub1) displayed restrained leaf and internode elongation, chlorophyll degradation, and carbohydrate consumption, whereas the enzymatic activities of pyruvate decarboxylase and alcohol dehydrogenase were increased significantly compared with the intolerant M202. Transcript levels of genes associated with carbohydrate consumption, ethanolic fermentation, and cell expansion were distinctly regulated in the two lines. Sub1A and Sub1C transcript levels were shown to be upregulated by submergence and ethylene, with the Sub1C allele in M202 also upregulated by treatment with gibberellic acid (GA). These findings demonstrate that the Sub1 region haplotype determines ethylene- and GA-mediated metabolic and developmental responses to submergence through differential expression of Sub1A and Sub1C. Submergence tolerance in lowland rice is conferred by a specific allele variant of Sub1A that dampens ethylene production and GA responsiveness, causing quiescence in growth that correlates with the capacity for regrowth upon desubmergence.

何云, 李贤伟, 李西, .

2种野生岩生植物叶片可溶性蛋白含量对低温胁迫的响应

安徽农业科学, 2008, 36(18):7552-7553.

[本文引用: 1]

齐光, 佟伟霜, 杨雨华, .

ABA对低温胁迫下水稻幼苗抗氧化酶活性的影响

湖北农业科学, 2016, 55(23):6079-6082.

[本文引用: 1]

刘丽, 甘志军, 王宪泽.

植物氮代谢硝酸还原酶水平调控机制的研究进展

西北植物学报, 2004, 24(7):1355-1361.

[本文引用: 1]

Martin A, Lee J, Kichey T, et al.

Two cytosolic glutamine synthetase isoforms of maize are specifically involved in the control of grain production

The Plant Cell, 2006, 18(11):3252-3274.

DOI:10.1105/tpc.106.042689      URL     [本文引用: 1]

The roles of two cytosolic maize glutamine synthetase isoenzymes (GS1), products of the Gln1-3 and Gln1-4 genes, were investigated by examining the impact of knockout mutations on kernel yield. In the gln1-3 and gln1-4 single mutants and the gln1-3 gln1-4 double mutant, GS mRNA expression was impaired, resulting in reduced GS1 protein and activity. The gln1-4 phenotype displayed reduced kernel size and gln1-3 reduced kernel number, with both phenotypes displayed in gln1-3 gln1-4. However, at maturity, shoot biomass production was not modified in either the single mutants or double mutants, suggesting a specific impact on grain production in both mutants. Asn increased in the leaves of the mutants during grain filling, indicating that it probably accumulates to circumvent ammonium buildup resulting from lower GS1 activity. Phloem sap analysis revealed that unlike Gln, Asn is not efficiently transported to developing kernels, apparently causing reduced kernel production. When Gln1-3 was overexpressed constitutively in leaves, kernel number increased by 30%, providing further evidence that GS1-3 plays a major role in kernel yield. Cytoimmunochemistry and in situ hybridization revealed that GS1-3 is present in mesophyll cells, whereas GS1-4 is specifically localized in the bundle sheath cells. The two GS1 isoenzymes play nonredundant roles with respect to their tissue-specific localization.

陶乐圆. 低温后施氮对水稻分蘖及氮代谢的影响. 哈尔滨:东北农业大学, 2018.

[本文引用: 2]

Arai-Sanoh Y, Ishimaru T, Ohsumi A, et al.

Effects of soil temperature on growth and root function in rice

Plant Production Science, 2010, 13(3):235-242.

DOI:10.1626/pps.13.235      URL     [本文引用: 1]

Makino A, Nakano H, Mae T.

Effects of growth temperature on the responses of ribulose-1,5-biphosphate carboxylase,electron transport components,and sucrose synthesis enzymes to leaf nitrogen in rice,and their relationships to photosynthesis

Plant Physiology, 1994, 105(4):1231-1238.

PMID:12232279      [本文引用: 1]

Effects of growth temperature on the photosynthetic gas-exchange rates and their underlying biochemical properties were examined in young, fully expanded leaves of rice (Oryza sativa L.). The plants were grown hydroponically under day/night temperature regimes of 18/15[deg]C, 23/18[deg]C, and 30/23[deg]C and all photosynthetic measurements were made at a leaf temperature of 25[deg]C and an irradiance of 1800 [mu]mol quanta m-2 s-1. Growth temperature affected the photosynthetic CO2 response curve. The relative ratio of the initial slope to the CO2-saturated photosynthesis increased with rising growth temperature. This was caused mainly by an increase in CO2-limited photosynthesis for a given leaf nitrogen content with rising growth temperature. However, there was no difference in ribulose-1,5-bisphosphate carboxylase (Rubisco) content at any given leaf nitrogen content among temperature treatments. In addition, the activation state and catalytic turnover rate of Rubisco were not affected by growth temperature. The increase in CO2-limited photosynthesis with rising growth temperature was the result of an increase in the CO2 transfer conductance between the intercellular airspaces and the carboxylation sites. The amounts of total chlorophyll and light-harvesting chlorophyll a/b protein II increased for the same leaf nitrogen content with rising growth temperature, but the amounts of cytochrome f and coupling factor 1 and the activities of cytosolic fructose-1,6-bisphosphatase and sucrose-phosphate synthase were the same between plants grown at 23/18[deg]C and those grown at 30/23[deg]C. Similarly, CO2-saturated photosynthesis was not different for the same leaf nitrogen content between these treatments. For the 18/15[deg]C-grown plants, a slight decrease in the amounts of cytochrome f and coupling factor 1 and an increase in the activities of cytosolic fructose-1,6-bisphosphatase and sucrose-phosphate synthase were found, but these were not reflected in CO2-saturated photosynthesis.

胡凝. 灌水对稻田微气象环境的影响及两系杂交稻安全制种的灌木技术. 南京:南京信息工程大学, 2005.

[本文引用: 1]

王柏, 黄彦, 孙艳玲, .

寒地高纬度区水稻水氮耦合效应试验研究

水利科学与寒区工程, 2020, 3(4):47-51.

[本文引用: 1]

徐国伟, 陆大克, 刘聪杰, .

干湿交替灌溉和施氮量对水稻内源激素及氮素利用的影响

农业工程学报, 2018, 34(7):137-146.

[本文引用: 1]

余小芬, 邱学礼, 张素华, .

低温与氮耦合对水稻内源激素含量的影响

西北农林科技大学学报, 2020, 48(9):73-80.

[本文引用: 1]

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