作物杂志, 2026, 42(1): 143-151 doi: 10.16035/j.issn.1001-7283.2026.01.018

生理生化·植物营养·栽培耕作

水稻叶片的生理性状和形态特征受铬胁迫的影响研究

刘晴,1, 孙露宏2, 高世伟1, 刘宇强1, 常汇琳1, 马成1, 王婧泽1, 王翠玲3, 聂守军,1

1黑龙江省农业科学院绥化分院,152000,黑龙江绥化

2农业农村部稻米及加工品质量与营养检验测试中心(桦川),154000,黑龙江佳木斯

3黑龙江省农业科学院农产品质量安全研究所,150000,黑龙江哈尔滨

Effects of Chromium Stress on Physiological Traits and Morphological Characteristics of Rice Leaves

Liu Qing,1, Sun Luhong2, Gao Shiwei1, Liu Yuqiang1, Chang Huilin1, Ma Cheng1, Wang Jingze1, Wang Cuiling3, Nie Shoujun,1

1Suihua Branch of Heilongjiang Academy of Agricultural Sciences, Suihua 152000, Heilongjiang, China

2Quality and Nutrition Inspection and Testing Centre for Rice and Processed Products (Huachuan), Ministry of Agriculture and Rural Affairs, Jiamusi 154000, Heilongjiang, China

3Quality and Safety Institute of Agricultural Products, Heilongjiang Academy of Agricultural Sciences, Harbin 150000, Heilongjiang, China

通讯作者: 聂守军,研究方向为水稻遗传育种,E-mail:nsj-0821@163.com

收稿日期: 2024-09-12   修回日期: 2024-11-9   网络出版日期: 2025-05-22

基金资助: 黑龙江省农业科技创新跨越工程农业科技基础创新优青项目(CX22YQ25)
黑龙江省农业科技创新跨越工程重大需求科技创新科技攻关项目(CX23ZD02)
黑龙江省水稻现代农业产业技术协同创新推广体系
黑龙江省农业科学院绥化分院科技创新项目(SHFY2022-01)
政府间国际科技创新合作(2022YFE0117800-4)
中国绿色食品发展中心农产品品质规格营养功能评价项目(GF-TSPZ-2024021)

Received: 2024-09-12   Revised: 2024-11-9   Online: 2025-05-22

作者简介 About authors

刘晴,研究方向为水稻遗传育种,E-mail:285394703@qq.com

摘要

以绥粳309(SJ309)和龙庆稻31(LQD31)2个水稻品种为材料,探究铬胁迫对水稻叶片生理性状与形态特征的影响。结果表明,铬主要在水稻根部富集,且SJ309比LQD31更有效地减少了铬的吸收与转移。在高浓度铬胁迫(100 μmol/L)下,2个品种的气孔导度、蒸腾速率、水分利用率及光合色素等生理特性均下降,叶片蒸气压亏缺、胞间CO₂浓度和丙二醛含量则升高。经铬胁迫处理,仅LQD31的气孔保卫细胞气孔面积增大、孔径闭合,而SJ309无明显变化。与SJ309相比,LQD31的脱落酸和水杨酸含量显著增加,导致参与气孔孔径调节的NCED1NCED2基因过度表达,表明LQD31对铬胁迫更为敏感。此外,铬胁迫使SJ309的非腺毛状体密度和长度显著增加,表明其能抵御紫外线损伤及多种环境胁迫。综上,水稻SJ309对铬胁迫耐受性更佳,其超积累特性可用于铬污染土壤的植物修复。

关键词: 铬胁迫; 水稻; 生理特性; 叶片; 形态特征

Abstract

Using two rice varieties, Suijing 309 (SJ309) and Longqingdao 31 (LQD31), as experimental materials, the effects of chromium (Cr) stress on the physiological traits and morphological characteristics of rice leaves were investigated. The results showed that Cr was primarily enriched in the roots of rice, and SJ309 was more effective than LQD31 in reducing the absorption and translocation of Cr. Under high-concentration Cr stress (100 μmol/L), the physiological characteristics such as stomatal conductance, transpiration rate, water use efficiency, and photosynthetic pigments decreased in both varieties, while leaf vapor pressure deficit, intercellular CO2 concentration, and malondialdehyde (MDA) content increased. After Cr stress treatment, only LQD31 exhibited an increase in the stomatal area of guard cells and closure of the stomatal aperture, whereas no significant changes were observed in SJ309. Compared with SJ309, the contents of abscisic acid (ABA) and salicylic acid (SA) in LQD31 increased significantly, leading to the overexpression of NCED1 and NCED2 genes involved in the regulation of stomatal aperture, which indicated that LQD31 was more sensitive to Cr stress. Furthermore, Cr stress significantly increased the density and length of non-glandular trichomes in SJ309, suggesting its ability to withstand UV damage and various environmental stresses. In summary, SJ309 demonstrates superior tolerance to Cr stress, and its hyperaccumulation characteristics can be utilized for the phytoremediation of Cr-contaminated soils.

Keywords: Chromium stress; Rice; Physiological characteristics; Leaf; Morphological characteristics

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本文引用格式

刘晴, 孙露宏, 高世伟, 刘宇强, 常汇琳, 马成, 王婧泽, 王翠玲, 聂守军. 水稻叶片的生理性状和形态特征受铬胁迫的影响研究. 作物杂志, 2026, 42(1): 143-151 doi:10.16035/j.issn.1001-7283.2026.01.018

Liu Qing, Sun Luhong, Gao Shiwei, Liu Yuqiang, Chang Huilin, Ma Cheng, Wang Jingze, Wang Cuiling, Nie Shoujun. Effects of Chromium Stress on Physiological Traits and Morphological Characteristics of Rice Leaves. Crops, 2026, 42(1): 143-151 doi:10.16035/j.issn.1001-7283.2026.01.018

铬是一种具有膜渗透性和高毒性的非必需重金属,可干扰组织细胞功能,甚至导致植物死亡[1-2],土壤铬污染还会造成各类农作物减产。铬在环境中主要以三价(Cr3+)和六价(Cr6+)离子形态存在[3]。铬酸盐和重铬酸盐化合物是铬的含氧阴离子,具有毒性、可溶于水且能释放游离的Cr6+,其来源大多为钢铁、电镀、皮革和化工等人为活动,而适量范围内的Cr3+并无毒性[4]。目前,植物吸收和转运铬的具体机制尚不明确。

重金属进入植物根表皮层主要是通过共质体与质外体途径进行的短程运输,以及木质部与韧皮部进行装卸的长程运输。植物对重金属的防御机制主要包括诱导产生金属硫蛋白、胁迫形成植物螯合肽及利用液泡对重金属进行区域化隔离[5]。植物通过细胞质膜吸收Cr6+,此过程涉及磷酸盐和硫酸盐转运体能量依赖性的活性转运[3,5]。研究[6-8]表明,较高浓度的Cr6+会对植物种子萌发、生长形态、生物量及产量产生不利影响。铬毒性会诱导植物产生活性氧(reactive oxygen species,ROS),如超氧阴离子(O2-. )、过氧化氢(H2O2)和羟自由基(·OH)等,进而干扰植物正常新陈代谢。ROS活性增强会破坏蛋白质、脂质和核酸等细胞结构。植物虽能通过产生超氧化物歧化酶(superoxide dismutase,SOD)、过氧化物酶(peroxidase,POD)和过氧化氢酶(catalase,CAT)等抗氧化酶来清除ROS,但在铬及其他重金属胁迫下,这些抗氧化酶的活性会降低[6,9-10]。在较高浓度铬胁迫下,小麦中丙二醛(malondialdehyde,MDA)浓度升高,这是由ROS活性增强导致的氧化损伤所致[11]

叶片生理性状与形态特征变化是植物对重金属胁迫作出的信息反馈。铬的植物毒性会减少实质细胞数量、改变叶绿体超微结构、抑制电子传递过程、缩小细胞间空间,并导致叶片气孔导度异常,进而使光合速率降低[12-13]。此外,铬还会干扰细胞的矿物质吸收,破坏水分平衡[6]。气孔的保卫细胞能对包括重金属胁迫在内的非生物胁迫作出快速响应,通过调节气孔的孔径和面积来适应环境变化。在非生物胁迫条件下,保卫细胞会积累更多脱落酸(abscisic acid,ABA),促使气孔关闭。尽管ABA和水杨酸(salicylic acid,SA)可能通过改变气孔的协同作用来调节植物对非生物胁迫的耐受性[14-15],但研究[16]指出,ABA依赖基因NCED1NCED2过表达会导致ABA积累,进而使气孔关闭、蒸腾速率下降。土壤中重金属浓度过高还会影响植物毛状体的结构完整性与分布。相关研究[17]显示,当砷浓度较高时,绿豆叶片正面和背面的毛状体密度均会降低。水稻叶片背面和正面的叶脉与表皮之间分布着腺毛状体和非腺毛状体,不过目前尚无研究报道水稻毛状体对土壤中较高浓度铬胁迫的反应情况。

水稻作为全球最重要的粮食作物之一,土壤重金属污染是其减产的主要原因之一。我国受重金属污染的土壤面积达上万公顷,占耕地总面积的15%以上,以每年数万吨粮食遭重金属污染计算,直接经济损失高达200亿元[18-19]。工业废品中含有的化学物质,是促使有毒金属在水稻植物组织中迁移的主要诱因[20-21]。然而,目前尚未见关于Cr6+毒性对水稻生理和形态影响的相关报道。本研究评估了Cr6+毒性对2个水稻品种叶片不同生理及形态特征参数的影响,旨在阐明高浓度Cr6+胁迫降低水稻叶片各项生理特性与形态特征参数的机制,为应对水稻重金属污染问题提供理论依据。

1 材料与方法

1.1 试验材料

供试材料为2个水稻品种绥粳309(SJ309)和龙庆稻31(LQD31),由黑龙江省农业科学院绥化分院提供。2个水稻品种均属于常规水稻,种植普及率较高,且具有高产、优质、多抗和耐旱等特点。

1.2 试验方法

选取外观健康且大小均匀的种子,于实验室可控环境生长室中繁育发芽,将发芽种子在18 h光照/6 h黑暗、30 °C/22 °C、相对湿度50%~60%以及光合有效辐射500±25 μmol/(m2·s)的环境条件下进行生长发育。2周后,分别用含有0(CK)、25(Cr25)、50(Cr50)和100 μmol/L(Cr100)Cr6+的重铬酸钾溶液处理2种供试水稻。由于经150 μmol/L铬胁迫处理20 d后的LQD31大部分已死亡,因此,最终确定试验所需的铬胁迫浓度范围在0~100 μmol/L。对照组和各浓度处理组均进行5次重复试验。在第3周至第5周,按上述相同条件每3 d更换1次处理液,共处理21 d。

1.3 测定指标与方法

1.3.1 铬(Cr6+)含量

经铬胁迫处理3周后,收获2个水稻品种的根和叶。于60 °C下干燥24 h后制样,称取0.5 g待测样品,使用7 mL HNO3进行微波消解,用水定容至50 mL。使用iCAPTM RQ ICP-MS电感耦合等离子体质谱仪(赛默飞世尔科技公司)测定Cr6+含量,绘制标准曲线,相关系数R2≥0.999。

1.3.2 生理性状与光合气体交换参数

使用CIRAS-3便携式光合作用测定系统(PP Systems,美国)分析CK和Cr100处理下2个水稻品种叶片的气孔导度(Gs)、蒸腾速率(Tr)、水分利用率(WUE)、胞间CO2浓度(Ci)、蒸气压亏缺和叶片温度。

1.3.3 叶片组织形态与生长指标

经铬胁迫处理3周后,在真空室中,用甲醛―乙醇―乙酸(formaldehyde-alcohol-acetic acid,FAA)固定叶片30 min,在初固定的15 min内需抽放真空数次。用碘化丙啶(10 μg/mL,磷酸盐缓冲液)对洗净的叶片染色15 min,随后放入甘油中。利用TCS SP8激光扫描共聚焦显微系统(徕卡,德国)在DMI600倒置显微镜(徕卡,德国)上运行,配备63×NA1.3甘油浸没式透镜用于成像,在采集透射光图像的同时使用488 nm激发波长采集荧光图像。利用ImageJ软件测定水稻茎长、根长和侧根数。定量分析时进行50次重复试验。

1.3.4 光合色素

分别称取CK与Cr100处理的2个水稻品种叶片样品(25 mg),用1.5 mL 80%(v/v)丙酮在4 ℃避光处理48 h。随后,将样品在13 000 g下离心10 min,将上清液稀释10倍后在663、645和480 nm波长下测定吸光度[22]。计算叶绿素和类胡萝卜素含量:叶绿素含量=(7.93×A663+ 19.53×A645)×f×V/FW,类胡萝卜素含量=(A480+ 0.114×A663-0.638×A645)×f×V/FW× 112.5,式中,f表示稀释因子,V表示样品溶液体积,FW表示收获待测样品的鲜重(mg)。

1.3.5 抗氧化酶活性与丙二醛(MDA)含量

采用硫代巴比妥酸(thiobarbituric acid,TBA)法测定MDA含量。称取0.2 g鲜叶样品,用5 mL含10%三氯乙酸的0.25% TBA溶液进行均质。在95 ℃下煮沸30 min,然后在10 000 g下离心10 min,于532 nm波长处测定吸光度,然后减去在600 nm波长处测得的吸光度(非特异性)。

参照Larkindale等[23]的方法,从100 mg叶片样品中提取抗氧化酶。用2 mL含有1%聚乙烯吡咯烷酮(PVP)和1 mmol/L乙二胺四乙酸(EDTA)的100 mmol/L磷酸盐缓冲液(pH=7.0)冷冻并研磨叶片样品。在4 ℃、13 000 g下离心20 min,将上清液分离并备用。

采用紫外分光光度法测定过氧化氢酶(CAT)活性,将0.1 mL酶提取物与1 mL 50 mmol/L磷酸钠缓冲液(pH=7.0)混匀,加入0.1 mL 100 mmol/L H2O2引发反应,每隔30s记录240 nm处的吸光度变化,持续2 min。采用愈创木酚法[24]测定过氧化物酶(POD)活性,将100 μL酶提取物与0.9 mL含有1 mmol/L EDTA的磷酸二氢钾缓冲液(pH=7.0)和10 mmol/L愈创木酚混合,加入0.1 mL 100 mmol/L H2O2引发反应,室温孵育5 min,于470 nm处记录吸光度变化。采用氮蓝四唑(NBT)光还原法测定超氧化物歧化酶(SOD)活性,将0.1 mL酶提取物加入189 μL 1 mmol/L硝基蓝四氮唑和39 μL 1 mmol/L甲硫氨酸中,再用50 mmol/L磷酸盐缓冲液(pH=7.8)定容,加入3.9 μL 1 mmol/L核黄素后,将反应混合物在15 W荧光灯下孵育10 min,于560 nm处测量吸光度。CAT、POD和SOD活性单位为U/(mg prot∙min)。

1.3.6 实时荧光定量PCR(qRT-PCR)

将CK和Cr100处理的2个水稻品种叶片用液氮速冻,并在-80°C保存备用。使用RNeasy Plant Mini Kit试剂盒(Qiagen,德国)提取总RNA。基于1 mg总RNA采用Reverse Transcription Supermix for RT-qPCR逆转录预混液(Bio-Rad,美国)获得cDNA。对于相对转录水平,使用CFX384 Touch™实时检测系统(Bio-Rad,美国),按照说明和iTaq Universal SYBR Green超混合液(Bio-Rad,美国),进行qRT-PCR的定量分析[25]。使用Primer Quest软件设计qRT- PCR中使用的基因特异性引物(表1),所有反应均在384孔PCR板中进行。使用肌动蛋白(内参基因)将转录水平归一化。每个qRT-PCR试验均进行3个重复。

表1   qRT-PCR引物信息

Table 1  Information of the primers for qRT-PCR

编号Code基因Gene正向引物Forward primer (5′→3′)反向引物Reverse primer (3′→5′)
1NCED1GCTCGGTCACTCACTCACTCGCGTTCTTCTTCCTGCCATAG
2NCED2CATGCTCCACTCCCTTCTCAGAAGCCAGCGAAGAAGTTTGG

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1.3.7 脱落酸(ABA)和水杨酸(SA)含量

参考Vadassery等[26]的方法,采用超高效液相色谱法对防御性植物激素(ABA和SA)进行测定与分析。将CK和Cr100处理的2个水稻品种叶片用液氮速冻,并在-80 ℃保存备用。叶片部分使用Tissuelyser-II组织研磨仪(上海净信实业发展有限公司)进行均质,于冻干机中保存过夜。取20 mg冻干样品用1 mL甲醇提取,甲醇中用40 ng/mL D6-ABA和40 ng/mL D4-SA作内标。将均质样品在振动器中混合30 min,然后在4 ℃下以13 000 g离心20 min。收集上清液,再次用500 μL甲醇提取匀浆并离心。合并提取物进行氮吹,并重新悬浮于500 μL甲醇中上机检测。

1.4 数据处理

采用方差分析(ANOVA)进行统计检验,使用Tukey检验进行事后多重比较,使用Origin 2021软件进行统计分析,显著性水平P<0.05,图表中的数据用平均值±标准差表示。

2 结果与分析

2.1 铬胁迫下水稻根和叶中的铬含量

水稻植株根和叶片中铬(Cr6+)含量的测定结果如表2所示,2个水稻品种根部铬积累量是叶的11.1~26.6倍,且随铬胁迫浓度升高,根和叶的铬积累量呈递增趋势。比较不同水稻品种根和叶中铬的富集量发现,LQD31的富集能力强于SJ309,在Cr100处理下差值最大。然而,CK处理下SJ309和LQD31的根和叶中均未检出铬,说明铬不易在水稻植株体内向茎部和叶片转移,且SJ309比LQD31能更有效地抑制植株对铬的吸收与转移。

表2   不同铬胁迫水平对水稻品种根和叶中Cr6+含量的影响

Table 2  Effects of different chromium stress levels on Cr6+ content in roots and leaves of rice varieties mg/g

品种Variety处理Treatment根Root叶Leaf
SJ309CK0.000±0.000e0.000±0.000e
Cr250.238±0.003d0.012±0.001d
Cr500.337±0.006c0.013±0.004d
Cr1000.452±0.009b0.017±0.003c
LQD31CK0.000±0.000e0.000±0.000e
Cr250.255±0.004d0.023±0.002b
Cr500.346±0.005c0.025±0.003b
Cr1000.550±0.011a0.034±0.005a

不同小写字母表示具有显著性差异(P < 0.05)。

Different lowercase letters indicate significant differences (P < 0.05).

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2.2 铬胁迫下水稻幼苗的生长指标

图1可知,2个水稻品种的茎长、根长和侧根数均有不同程度的降低(图1)。Cr100处理下水稻植株的茎长、根长和侧根数均极显著减少,与SJ309相比LQD31的降幅较大,受铬胁迫影响更明显。然而,与CK处理相比,Cr25处理下2个水稻品种的茎长无明显变化,只有在Cr100处理下SJ309和LQD31的茎长受铬胁迫影响极显著,分别下降了24.2%和22.1%。

图1

图1   不同铬胁迫水平对水稻品种幼苗生长指标的影响

“***”表示处理间差异达极显著水平(P < 0.001),下同。

Fig.1   Effects of different chromium stress levels on seedling growth index of rice varieties

“***”indicates extremely significant difference among treatments at P < 0.001 level, the same below.


2.3 铬胁迫下水稻叶片的光合气体交换参数和生理性状

Cr100处理下,水稻植株的Gs图2a)、Tr图2b)和WUE图2c)均有所下降,蒸气压亏缺(图2d)则有所上升。Cr100处理下SJ309和LQD31的叶片温度分别提高了6.3%和5.1%,表明铬胁迫下水稻叶温在不同品种之间无显著差异(图2e)。由图2f可知,铬胁迫对Ci具有一定的影响,SJ309的Ci减少4.9%,LQD31则增加19.4%。Cr100处理下LQD31的Ci增加导致GsTr降低。较高的蒸气压亏缺导致LQD31植株的GsTrWUE显著下降。

图2

图2   铬胁迫对不同水稻品种光合气体交换参数和生理性状的影响

“**”表示处理间差异达极显著水平(P < 0.01),下同。

Fig.2   Effects of chromium stress on photosynthetic gas exchange parameters and physiological traits of different rice varieties

“**”indicates extremely significant difference among treatments (P < 0.01), the same below.


2.4 铬胁迫下水稻叶片的组织形态

图3a可知,CK处理下SJ309和LQD31叶片中的气孔保卫细胞为正常形态,Cr100处理下SJ309叶片上的气孔保卫细胞保持正常形态,但LQD31的保卫细胞则出现变形。由图3c图3d可知,SJ309和LQD31的气孔面积均有所增加,增幅分别为5.0%和30.8%。CK处理下2个水稻品种叶片和Cr100处理下SJ309叶片的气孔孔径均为开放状态,但Cr100处理下LQD31叶片的气孔孔径关闭,SJ309在CK和Cr100处理下的气孔孔径面积无显著差异。由图3b图3e可知,Cr100处理下SJ309和LQD31的气孔密度均有所增加,但不显著。

图3

图3   铬胁迫对不同水稻品种叶片气孔形态的影响

Fig.3   Effects of chromium stress on leaf stomatal morphology of different rice varieties


图4a可知,CK和Cr100处理下的水稻植株叶片上可观察到2种毛状体,即腺毛状体(腺毛,绿圈)和非腺毛状体(非腺毛,黄圈)。由图4b可知,CK和Cr100处理下2个水稻品种叶片的腺毛和非腺毛密度有所不同,铬胁迫对LQD31的腺毛和SJ309的非腺毛影响较为显著,增幅分别为80.3%和183.7%。铬胁迫对不同品种非腺毛密度的影响结果相反,SJ309的非腺毛密度增大,LQD31的非腺毛密度则下降11.9%。SJ309和LQD31的腺毛长度在不同铬处理间无显著差异。在非腺毛状体长度方面(图4c),Cr100处理下SJ309增加了37.2%,LQD31则减少了26.1%。

图4

图4   铬胁迫对不同水稻品种叶片毛状体形态的影响

Fig.4   Effects of chromium stress on leaf trichome morphology of different rice varieties


2.5 铬胁迫下水稻的光合色素与抗氧化酶活性

水稻的光合色素含量受铬胁迫的影响(图5a~b),Cr100处理下SJ309和LQD31叶片中叶绿素含量分别减少了22.0%和65.8%,类胡萝卜素含量分别减少了8.1%和65.2%,表明LQD31叶片受铬胁迫的影响更为显著。Cr100处理下,SJ309的CAT活性降低了21.0%,LQD31则提高了16.3%(图5c);SJ309和LQD31的POD活性分别提高了3.0%和34.9%(图5d);SJ309的SOD活性降低了9.1%,LQD31则提高了7.0%(图5e);SJ309和LQD31的MDA含量分别提高了1.0%和34.2%,LQD31材料Cr100与CK处理相比有显著差异(图5f)。

图5

图5   铬胁迫对不同水稻品种光合色素与抗氧化酶活性的影响

Fig.5   Effects of chromium stress on photosynthetic pigments and antioxidant enzyme activities in different rice varieties


2.6 铬胁迫下水稻植物激素的调控与表达

基于CK与Cr100处理植物激素调控与表达的差异分析(图6),探究不同水稻品种气孔保卫细胞孔径的分子调控作用。Cr100处理下,SJ309和LQD31叶片中NCED1基因的表达水平分别下调了72.3%和上调了315.1%,NCED2基因的表达水平分别下调了46.2%和上调了280.8%。SJ309和LQD31叶片中ABA含量分别增加了7.2%和30.7%,SA含量则分别为减少8.1%和增加43.0%。综上,铬胁迫下LQD31中ABA依赖的NCED1NCED2基因水平上调,同时ABA和SA的协同作用增强,共同参与调控水稻叶片气孔孔径的关闭过程。

图6

图6   铬胁迫对不同水稻品种植物激素调控与表达的影响

Fig.6   Effects of chromium stress on regulation and expression of plant hormones in different rice varieties


3 讨论

由水稻LQD31根和叶中铬富集量较高以及各项生理参数的检测结果可知,LQD31因无法对铬毒性进行螯合与解离,相较于SJ309更易在根部积累Cr6+,并经茎部向叶片转移。Daud等[8]和Singh等[27]针对棉花和绿豆品种的研究均报道了类似对铬胁迫的反应。2个水稻品种SJ309和LQD31的根部均积累了较高浓度的Cr6+,这是由于Cr6+固定于根细胞的液泡中,此为植物应对毒性的自然反应。由于根系是重金属元素的首要接触部位,根部的重金属积累量高于茎和叶[28-29]。铬胁迫诱导的植物毒性效应表现为2个水稻品种生长参数下降。Jabeen等[30]研究表明,铬是毒性最强的金属,会影响农作物植株的根长和侧根数,本研究结果与其一致。根长缩短是由于根细胞壁的细胞分裂受到抑制,茎长缩短则归因于叶片细胞的超微结构变形。

在铬胁迫条件下,LQD31的GsTrWUE显著降低,证实了不同水稻品种对铬胁迫存在特定响应。Vernay等[13]研究发现黑麦草植株在较高浓度的铬胁迫下,其GsTrWUE下降,同时Ci升高,本研究结果与其相似。光合作用参数的降低,可能是由于Cr6+穿透细胞膜引发叶绿素分解所致,此外,气孔对蒸气压亏缺和Ci的响应特性在物种内存在差异[6,31],这一结论与本研究结果相符。

较高浓度的CO2和蒸气压亏缺会促使气孔关闭,这可能与LQD31在气压不足、CO2浓度升高时气孔保卫细胞孔径关闭有关。经铬胁迫处理后,水稻叶片气孔面积增大,表明铬及其他重金属的同化与沉积会干扰气孔的功能形态,进而影响整体的光合气体交换参数,这与Chandra等[32]的结果一致。铬胁迫下LQD31的保卫细胞变形,Chen等[33]研究发现这可能是由于气孔保卫细胞细胞壁的含水量和机械性能改变,引发膨压增大所致,研究[34]中也观察到了类似的气孔保卫细胞变形现象。铬毒性导致水稻植株叶片缺水,引发SJ309和LQD31气孔密度增大。植物接触铬后,毛状体数量会发生变化,而毛状体可能是金属元素的储存区域。LQD31腺毛密度增加,表明该品种水稻是植物精油以及各类挥发性和非挥发性次级代谢产物的来源,这些代谢产物可为植物抵御食草动物提供保护[35]。重金属胁迫会影响水稻腺毛状体的形态和超微结构。此外,铬毒性还会导致SJ309非腺毛状体密度和长度增加,表明SJ309既能抵御紫外线造成的损伤、稳定温度,又能作为抵御食草动物的屏障,同时也说明SJ309比LQD31更能耐受多种环境胁迫。

经Cr6+胁迫处理后,SJ309水稻叶片中的SOD和CAT活性降低,这一结果与Tang等[36]的研究一致,证实氧化胁迫可能导致这2种抗氧化酶活性下降。然而,POD在叶片组织中的定位可能促使水稻POD活性增强[37]。MDA含量是反映ROS对植物膜造成损伤的重要指标之一。与SJ309相比,LQD31的MDA含量更高,表明其叶片细胞膜和超微结构可能因铬毒性而受损,Basit等[10]研究发现这可能是由铬毒性引发的氧化应激导致ROS活性增加所致。2个水稻品种中均发现ROS产生的氧化应激,但LQD31受影响更为显著,表明SJ309能更有效地消除ROS活性并抵御铬胁迫。Cr100处理下,2个品种的叶绿素和类胡萝卜素含量均下降,这与ROS的氧化应激有关,且LQD31的光合色素受影响程度更为显著。

铬胁迫会导致依赖ABA的NCED1NCED2基因过度表达。经铬胁迫处理的LQD31叶片,其Gs下降且气孔关闭,铝胁迫下的柑橘也呈现出类似现象[38]。相关研究[16,39]表明,非生物胁迫下NCED1NCED2基因的过度表达与蒸气压亏缺的增加有关,与本研究结果一致。NCED1NCED2基因过量表达会促使更多ABA积累,进而降低GsTr,最终导致气孔关闭。Zeng等[40]研究发现保卫细胞中的SA和ABA信号具有协同作用,在多种非生物胁迫下可协同调节气孔孔径大小。经铬胁迫处理的LQD31中SA和ABA含量增加,Prodhan等[15]研究表明这些植物激素在气孔关闭过程中协同传递信号。

本研究不仅探究了水稻应对Cr6+胁迫的生理性状与形态特征适应机制,还提出了一种依据多种生理及形态特征参数筛选抗Cr6+胁迫水稻品种的方法。基于在SJ309和LQD31中观察到的生理、形态及生化参数变化,不同水稻品种对Cr6+毒性的耐受能力存在差异。结果表明,SJ309已具备多种对Cr6+毒性的耐受机制,可被视为一种超积累水稻植株,可通过不同的基因表达将Cr6+毒性螯合并解离至液泡中[27,41]。深入探究SJ309和LQD31中参与Cr6+积累、转运和解毒机制的分子途径,将为作物育种者在铬污染土壤中选择和培育水稻栽培品种提供更优的方案。SJ309的超积累特性可用于铬污染土壤的植物修复,因此,作物育种者可采用轮作策略以提高这2个水稻品种在铬污染土壤中的产量。

4 结论

铬胁迫下,2个水稻品种植株的茎长、根长和侧根数均极显著减少,铬主要富集于根部,且SJ309比LQD31更有效减少铬的吸收与转移。二者气孔面积和密度均增加,LQD31变化更明显,其气孔保卫细胞变形、孔径闭合;铬胁迫对SJ309毛状体形态影响较大。高浓度铬胁迫(Cr100)下,2个品种的气孔导度、蒸腾速率、水分利用率、光合色素含量、CAT和SOD活性降低,蒸气压亏缺、胞间CO₂浓度、MDA含量和POD活性升高,LQD31变化幅度更大,对铬胁迫更敏感。ABA和SA含量增加促使NCED1NCED2基因过度表达,导致LQD31叶片气孔导度降低、孔径关闭。水稻SJ309对铬胁迫耐受性更好,其超积累特性可用于铬污染土壤的植物修复。

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不同栽培条件对水稻籽粒镉含量及主要性状的影响研究

作物杂志, 2024(4):203-208.

[本文引用: 1]

Ochoa M, Tierra W, Tupuna-Yerovi D S, et al.

Assessment of cadmium and lead contamination in rice farming soils and rice (Oryza sativa L.)

from Guayas province in Ecuador. Environmental Pollution, 2020,260:114050.

[本文引用: 1]

Geider R J, Osborne B A. Algal Photosynthesis. Boston:Springer,1992.

[本文引用: 1]

Larkindale J, Huang B R.

Thermotolerance and antioxidant systems in Agrostis stolonifera: involvement of salicylic acid, abscisic acid, calcium, hydrogen peroxide, and ethylene

Journal of Plant Physiology, 2004, 161(4):405-413.

PMID:15128028      [本文引用: 1]

This study investigated whether pre-treating plants with specific putative signaling components and heat acclimation would induce tolerance of a cool-season grass, creeping bentgrass (Agrostis stolonifera var. palustris), to subsequent heat stress and whether thermotolerance induction of those pretreatments was associated with the regulation of antioxidant regenerating enzymes. The treatments included foliar application of salicylic acid (SA), abscisic acid (ABA), calcium chloride (CaCl2), hydrogen peroxide (H2O2), 1-aminocyclopropane-1-carboxylic acid (ACC, a precursor of ethylene prior to the exposure of plants to heat stress (35 degrees C) in a growth chamber. Physiological measurements including turf quality, leaf photosynthetic rate, and levels of oxidative damage demonstrated that all treatments increased heat tolerance. The better heat tolerance for pre-treated plants as compared to controls was related to the protection of oxidative damage under heat stress. APX activity increased over the first 2 days and 5 days of heating for ACC and CaCl2 respectively, but for only 12 h for H2O2. SA and ABA pre-treatments had no effects on APX activity earlier, but maintained APX activity at a significantly higher level than in controls after 24 h of heating. SA and ABA pre-treatments had no effects on POX activity. ACC treatment significantly increased POX activity. Pre-treatment with CaCl2, H2O2, and HA reduced POX activity, particularly during the later phase of heating. Plants treated with SA, CaCl2, H2O2 and HA had lower CAT activity than their control plants prior to heating and within 48 h of heat stress. ABA and ACC pre-treatments maintained higher CAT activity than the controls after 48 h of heating. ACC, CaCl2, or HA pre-treatments increased SOD activity only before 5 days of heat stress. SA and ABA pre-treatments had less effect on APX activity earlier under heat stress. These results suggest that specific groups of potential signaling molecules may induce tolerance of creeping bentgrass to heat stress by reducing oxidative damage.

Saha B, Mishra S, Awasthi J P, et al.

Enhanced drought and salinity tolerance in transgenic mustard [Brassica juncea (L.) Czern & Coss.] overexpressing Arabidopsis group 4 late embryogenesis abundant gene (AtLEA4-1)

Environmental and Experimental Botany, 2016,128:99-111.

[本文引用: 1]

Kumar A, Panigrahy M, Panigrahi K C.

Optimization of soil parameters and cost effective way of growing Arabidopsis thaliana from an Indian perspective

International Journal of Basic and Applied Agricultural Research, 2018, 16(1):54-59.

[本文引用: 1]

Vadassery J, Reichelt M, Hause B, et al.

CML42-mediated calcium signaling coordinates responses to Spodoptera herbivory and abiotic stresses in Arabidopsis

Plant Physiology, 2012, 159 (3):1159-1175.

DOI:10.1104/pp.112.198150      URL     [本文引用: 1]

In the interaction between Arabidopsis (Arabidopsis thaliana) and the generalist herbivorous insect Spodoptera littoralis, little is known about early events in defense signaling and their link to downstream phytohormone pathways. S. littoralis oral secretions induced both Ca2+ and phytohormone elevation in Arabidopsis. Plant gene expression induced by oral secretions revealed up-regulation of a gene encoding a calmodulin-like protein, CML42. Functional analysis of cml42 plants revealed more resistance to herbivory than in the wild type, because caterpillars gain less weight on the mutant, indicating that CML42 negatively regulates plant defense; cml42 also showed increased aliphatic glucosinolate content and hyperactivated transcript accumulation of the jasmonic acid (JA)-responsive genes VSP2 and Thi2.1 upon herbivory, which might contribute to increased resistance. CML42 up-regulation is negatively regulated by the jasmonate receptor Coronatine Insensitive1 (COI1), as loss of functional COI1 resulted in prolonged CML42 activation. CML42 thus acts as a negative regulator of plant defense by decreasing COI1-mediated JA sensitivity and the expression of JA-responsive genes and is independent of herbivory-induced JA biosynthesis. JA-induced Ca2+ elevation and root growth inhibition were more sensitive in cml42, also indicating higher JA perception. Our results indicate that CML42 acts as a crucial signaling component connecting Ca2+ and JA signaling. CML42 is localized to cytosol and nucleus. CML42 is also involved in abiotic stress responses, as kaempferol glycosides were down-regulated in cml42, and impaired in ultraviolet B resistance. Under drought stress, the level of abscisic acid accumulation was higher in cml42 plants. Thus, CML42 might serve as a Ca2+ sensor having multiple functions in insect herbivory defense and abiotic stress responses.

Singh S, Parihar P, Singh R, et al.

Heavy metal tolerance in plants: role of transcriptomics, proteomics, metabolomics, and ionomics

Frontiers in Plant Science, 2016,6:165395.

[本文引用: 2]

Shanker A K, Djanaguiraman M, Sudhagar R, et al.

Differential antioxidative response of ascorbate glutathione pathway enzymes and metabolites to chromium speciation stress in green gram (Vigna radiata (L.) R. Wilczek. cv CO 4) roots

Plant Science, 2004, 166(4):1035-1043.

DOI:10.1016/j.plantsci.2003.12.015      URL     [本文引用: 1]

Feleafel M N, Mirdad Z M.

Hazard and effects of pollution by lead on vegetable crops

Journal of Agricultural and Environmental Ethics, 2013, 26(3):547-567.

DOI:10.1007/s10806-012-9403-1      URL     [本文引用: 1]

Jabeen N, Abbas Z, Iqbal M, et al.

Glycinebetaine mediates chromium tolerance in mung bean through lowering of Cr uptake and improved antioxidant system

Archives of Agronomy and Soil Science, 2016, 62(5):648-662.

DOI:10.1080/03650340.2015.1082032      URL     [本文引用: 1]

Atta M I, Bokhari T Z, Malik S A, et al.

Assessing some emerging effects of hexavalent chromium on leaf physiological performance in sunflower (Helianthus annuus L.)

International Journal of Scientific & Engineering Research, 2013, 4(8):945-949.

[本文引用: 1]

Chandra R, Kang H.

Mixed heavy metal stress on photosynthesis, transpiration rate, and chlorophyll content in poplar hybrids

Forest Science and Technology, 2016, 12(2):55-61.

DOI:10.1080/21580103.2015.1044024      URL     [本文引用: 1]

Chen Y T, Li W L, Turner J A, et al.

PECTATE LYASE LIKE 12 patterns the guard cell wall to coordinate turgor pressure and wall mechanics for proper stomatal function in Arabidopsis

The Plant Cell, 2021, 33(9):3134-3150.

DOI:10.1093/plcell/koab161      URL     [本文引用: 1]

Plant cell deformations are driven by cell pressurization and mechanical constraints imposed by the nanoscale architecture of the cell wall, but how these factors are controlled at the genetic and molecular levels to achieve different types of cell deformation is unclear. Here, we used stomatal guard cells to investigate the influences of wall mechanics and turgor pressure on cell deformation and demonstrate that the expression of the pectin-modifying gene PECTATE LYASE LIKE12 (PLL12) is required for normal stomatal dynamics in Arabidopsis thaliana. Using nanoindentation and finite element modeling to simultaneously measure wall modulus and turgor pressure, we found that both values undergo dynamic changes during induced stomatal opening and closure. PLL12 is required for guard cells to maintain normal wall modulus and turgor pressure during stomatal responses to light and to tune the levels of calcium crosslinked pectin in guard cell walls. Guard cell-specific knockdown of PLL12 caused defects in stomatal responses and reduced leaf growth, which were associated with lower cell proliferation but normal cell expansion. Together, these results force us to revise our view of how wall-modifying genes modulate wall mechanics and cell pressurization to accomplish the dynamic cellular deformations that underlie stomatal function and tissue growth in plants.

Gautam V, Kohli S K, Kapoor D, et al.

Stress protective effect of Rhododendron arboreum leaves (MEL) on chromium-treated Vigna radiata plants

Journal of Plant Growth Regulation, 2021, 40(1):423-435.

DOI:10.1007/s00344-020-10111-6      [本文引用: 1]

Khetnon P, Busarakam K, Sukhaket W, et al.

Mechanisms of trichomes and terpene compounds in indigenous and commercial Thai rice varieties against brown planthopper

Insects, 2022, 13 (5):427.

DOI:10.3390/insects13050427      URL     [本文引用: 1]

Plant trichomes generally act as a physical defense against herbivore attacks and are present in a variety of plants, including rice plants. This research examined the physical and chemical defenses of rice plants against the brown planthopper (BPH), Nilaparvata lugens (Stål) (Hemiptera: Delphacidae). A total of 10 rice varieties were used in this study. An electron microscope was used to observe trichomes. Constitutive and induced volatile compound profiles were assessed using GC-MS analyses. The preference of BPH for volatiles from the 10 rice plants was tested using a two-choice arena olfactometer system. The density of prickle trichomes had a negative relationship with the BPH injury level. Without BPH infestation, the volatile of the most resistant rice variety (Rathu Heenati (RH)) was preferred by BPH than those of the other varieties, with the exception of Gled Plah Chawn. However, the relative BPH preference for volatiles from the RH variety decreased during BPH infestation. When rice plants were infested by BPH, the numbers of VOCs and these quantities decreased. In the RH variety, the emission of essentities found without BPH infestation ceased during infestation by BPH. During the BPH infestation, rice plants started to emit new VOCs that were not detected before the BPH infestation started. In conclusion, we discovered that rice plants defended against BPH by changing VOC components during BPH infestation and β-Sesquiphellandrene was likely the most effective component.

Tang S, Liu Y L, Zheng N, et al.

Temporal variation in nutrient requirements of tea (Camellia sinensis) in China based on QUEFTS analysis

Scientific Reports, 2020, 10(1):1745.

DOI:10.1038/s41598-020-57809-x      PMID:32019970      [本文引用: 1]

Fertilisation datasets collected from field experiments (n = 21) in tea-producing areas from 2016 to 2018 were used to build a quantitative evaluation of the fertility of tropical soils (QUEFTS) model to estimate nutrient uptake of tea plants, and to investigate relationships between tea yield and nutrient accumulation. The production of 1000 kg spring tea (based on one bud with two young expanding leaves) required 12.2 kg nitrogen (N), 1.2 kg phosphorus (P), and 3.9 kg potassium (K), and the corresponding internal efficiencies (IEs) for N, P, and K were 82.0, 833.3, and 256.4 kg kg. To produce 1000 kg summer tea, 9.1 kg N, 0.8 kg P, and 3.1 kg K were required, and the corresponding IEs for N, P, and K were 109.9, 1250.0, and 322.6 kg kg. For autumn tea, 8.8 kg N, 1.0 kg P, and 3.2 kg K were required to produce 1000 kg tea, and the corresponding IEs for N, P, and K were 113.6, 1000.0, and 312.5 kg kg. Field validation experiments performed in 2019 suggested that the QUEFTS model can appropriately estimate nutrient uptake of tea plants at a certain yield and contribute to developing a fertiliser recommendation strategy for tea production.

Zaheer I E, Ali S, Saleem M H, et al.

Role of iron-lysine on morpho-physiological traits and combating chromium toxicity in rapeseed (Brassica napus L.) plants irrigated with different levels of tannery wastewater

Plant Physiology and Biochemistry, 2020,155:70-84.

[本文引用: 1]

Gavassi M A, Silva G S, da Silva C M S, et al.

NCED expression is related to increased ABA biosynthesis and stomatal closure under aluminum stress

Environmental and Experimental Botany, 2021,185:104404.

[本文引用: 1]

Speirs J, Binney A, Collins M, et al.

Expression of ABA synthesis and metabolism genes under different irrigation strategies and atmospheric VPDs is associated with stomatal conductance in grapevine (Vitis vinifera L. cv Cabernet Sauvignon)

Journal of Experimental Botany, 2013, 64(7):1907-1916.

DOI:10.1093/jxb/ert052      PMID:23630325      [本文引用: 1]

The influence of different levels of irrigation and of variation in atmospheric vapour pressure deficit (VPD) on the synthesis, metabolism, and transport of abscisic acid (ABA) and the effects on stomatal conductance were examined in field-grown Cabernet Sauvignon grapevines. Xylem sap, leaf tissue, and root tissue were collected at regular intervals during two seasons in conjunction with measurements of leaf water potential (Ψleaf) and stomatal conductance (gs). The different irrigation levels significantly altered the Ψleaf and gs of the vines across both seasons. ABA abundance in the xylem sap was correlated with gs. The expression of genes associated with ABA synthesis, NCED1 and NCED2, was higher in the roots than in the leaves throughout and highest in the roots in mid January, a time when soil moisture declined and VPD was at its highest. Their expression in roots was also inversely related to the levels of irrigation and correlated with ABA abundance in the roots, xylem sap, and leaves. Three genes encoding ABA 8'-hydroxylases were isolated and their identities confirmed by expression in yeast cells. The expression of one of these, Hyd1, was elevated in leaves when VPD was below 2.0-2.5 kPa and minimal at higher VPD levels. The results provide evidence that ABA plays an important role in linking stomatal response to soil moisture status and that changes in ABA catabolism at or near its site of action allows optimization of gas exchange to current environmental conditions.

Zeng W, Melotto M, He S Y.

Plant stomata: a checkpoint of host immunity and pathogen virulence

Current Opinion in Biotechnology, 2010, 21(5):599-603.

DOI:10.1016/j.copbio.2010.05.006      PMID:20573499      [本文引用: 1]

Stomata are microscopic pores formed by pairs of guard cells in the epidermis of terrestrial plants; they are essential for gas exchange with the environment and controlling water loss. Accordingly, plants regulate stomatal aperture in response to environmental conditions, such as relative humidity, CO(2) concentration, and light intensity. Stomatal openings are also a major route of pathogen entry into the plant and plants have evolved mechanisms to regulate stomatal aperture as an immune response against bacterial invasion. In this review, we highlight studies that begin to elucidate signaling events involved in bacterium-triggered stomatal closure and discuss how pathogens may have exploited environmental conditions or, in some cases, have evolved virulence factors to actively counter stomatal closure to facilitate invasion.Copyright © 2010 Elsevier Ltd. All rights reserved.

Elango D, Devi K D, Jeyabalakrishnan H K, et al.

Agronomic, breeding, and biotechnological interventions to mitigate heavy metal toxicity problems in agriculture

Journal of Agriculture and Food Research, 2022,10:100374.

[本文引用: 1]

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