甜菜种质资源苗期耐旱性综合评价
Comprehensive Evaluation of Drought Tolerance of Sugar Beet Germplasms at Seedling Stage
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收稿日期: 2022-04-21 修回日期: 2022-06-30 网络出版日期: 2022-10-09
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Received: 2022-04-21 Revised: 2022-06-30 Online: 2022-10-09
作者简介 About authors
李王胜,研究方向为甜菜种质资源鉴定,E-mail:
对336份甜菜种质资源苗期的15个表型和生理生化指标进行测定,并通过隶属函数、主成分分析、聚类分析和相关性分析方法对甜菜种质资源耐旱性进行综合评价。结果表明,干旱胁迫后叶干重、株高、根长、叶鲜重、根鲜重、根干重、叶片饱和鲜重和叶片相对含水量等指标均显著降低,根冠比、可溶性糖、可溶性蛋白和脯氨酸含量等4个指标均显著高于对照。主成分分析将15个单一指标转化为6个综合指标,可代表原始数据信息的75.95%。聚类分析将336份甜菜种质资源分为5个类群,其中耐旱性强种质16份,耐旱性较强种质49份,耐旱性中等种质109份,耐旱性较弱种质79份,耐旱性弱种质83份。相关性分析结果显示,胚轴直径、株高、根长、叶鲜重、根鲜重、根干重、叶干重、叶片饱和鲜重、叶片相对含水量和根冠比与D值呈显著相关性。
关键词:
In this study, 336 sugar beet germplasm resources were measured at the seedling stage along with 15 phenotypic, physiological and biochemical indices, and the drought tolerance of these resources was thoroughly assessed using the affiliation function method, principal component analysis, cluster analysis and correlation analysis. The results showed that the indicators of leaf dry weight, plant height, root length, leaf fresh weight, root fresh weight, root dry weight, leaf saturated fresh weight and leaf relative water content were significantly decreased, and root to crown ratio, soluble sugar content, soluble protein content and proline content under drought stress were significantly higher than the control. The principal component analysis transformed the 15 single indicators into six composite indicators that could represent 75.95% of the original data information. Cluster analysis divided the 336 sugar beet germplasm resources into five groups, including 16 germplasms with strong drought tolerance, 49 with relatively strong drought tolerance, 109 with medium drought tolerance, 79 with relatively weak drought tolerance and 83 with weak drought tolerance. Correlation analysis showed that embryonic axis diameter, plant height, root length, leaf fresh weight, root fresh weight, root dry weight, leaf dry weight, leaf saturated fresh weight, relative leaf water content and root-shoot ratio were significantly correlated with D values.
Keywords:
本文引用格式
李王胜, 王雪倩, 尹希龙, 石杨, 刘大丽, 谭文勃, 兴旺.
Li Wangsheng, Wang Xueqian, Yin Xilong, Shi Yang, Liu Dali, Tan Wenbo, Xing Wang.
甜菜是世界上重要的糖料作物,也可作为饲料原料[1],随着对甜菜的不断研究,目前发现甜菜具有保健功能,如在降脂、降压等方面[2-3],应用前景十分广泛。种质资源又称遗传资源,中国保存的种质资源总量突破52万份,位居世界第二[4],我国国家甜菜种质中期库现保存甜菜种质资源1382份[5],种质资源作为农业“芯片”在科学研究中越来越受重视。我国甜菜主要种植在新疆、内蒙古和黑龙江等地[6],这些地区的年降水量多在400mm以下[7],并且近年来全球变暖造成水资源短缺,干旱成为影响我国甜菜生产的主要非生物因素[8]。苗期耐旱能力直接影响甜菜的生长以及产量,因此对甜菜种质资源进行苗期耐旱试验,筛选出优异的耐旱种质资源,对促进我国农业发展、加强生态环境安全和可持续发展具有重要意义。
本研究以336份国家甜菜种质中期库中保存的甜菜种质资源为材料,通过PEG-6000模拟干旱胁迫对甜菜苗期的株高、根长、可溶性糖和游离脯氨酸含量等15个指标进行分析。通过隶属函数法、主成分分析法、相关分析法和聚类分析法综合评价甜菜种质资源的耐旱性,试验主要目的是进一步完善甜菜种质资源耐旱评价体系和评价指标,筛选出苗期耐旱种质资源,为今后甜菜种质资源耐旱研究以及育种提供材料。
1 材料与方法
1.1 试验材料
本研究以国家甜菜种质中期库提供的来源于17个国家的336份甜菜种质资源为材料,其中中国205份、英国2份、意大利5份、匈牙利2份、瑞典5份、日本9份、俄罗斯16份、美国49份、罗马尼亚2份、荷兰12份、法国3份、德国10份、丹麦1份、朝鲜1份、波兰11份、比利时2份以及奥地利1份。
1.2 试验设计
试验于2021年3-12月在黑龙江大学国家甜菜种质中期库(126°45′ E,45°30′ N)智能人工气候室内进行,以PEG-6000模拟干旱胁迫,通过实验室前期试验设置9%浓度时胁迫效果显著且未致死,所以确定设置干旱9% PEG-6000的Hoagland溶液(DS)和对照Hoagland溶液(CK)2个处理组,重复3次。
每份种质挑选30颗籽粒饱满、大小均匀的种子,用75%酒精浸泡1min后用自来水冲洗1min,在2‰的福美双溶液浸泡12h,将种子表面残留药物用自来水冲洗干净并播于蛭石中,每天定时喷水,待子叶长出,每份挑选长势一致的幼苗12株,洗去根上的蛭石后用海绵定植于24孔水培盆(长37.5cm,宽25.5cm,高13.5cm)中,盆中加入Hoagland营养液9L,每7d更换1次营养液,定植14d后(3对真叶时)进行干旱和对照处理,通过实验室前期试验设置胁迫2、5和7d,7d时效果最好,所以确定胁迫7d后测定最大根长、株高、根鲜重、叶鲜重、叶片饱和鲜重、叶干重、根干重、胚轴直径、叶片相对含水量、根冠比、超氧化物歧化酶(superoxide dismutase,SOD)活性以及可溶性糖、可溶性蛋白、游离脯氨酸和丙二醛含量。
1.3 测定项目与方法
最大根长(root length,RL)和株高(plant height,PH)用直尺测量,根鲜重(root fresh weight,RFW)、叶鲜重(leaf fresh weight,LFW)、叶片饱和鲜重(leaf saturated fresh weight,LSFW)、叶干重(leaf dry weight,LDW)和根干重(root dry weight,RDW)用电子天平称重,胚轴直径(embryonic axis diameter,EAD)使用游标卡尺测量。
叶片相对含水量(relative leaf water,RLW):RLW[16]=(LWF-LDW)/(LSFW-LDW)
根冠比(root-shoot ratio,RSR):RSR=RFW/ LFW
1.4 数据处理
分析中涉及的计算公式如下:
式中,Xi为第i个指标干旱处理后与对照处理的相对值,Xi min和Xi max分别表示第i个指标耐旱系数的最小值和最大值。
式中,F(Xj)表示第j个综合指标值,aij表示各指标所对应的特征值,Xij为各指标标准化处理的值。
式中,Wj表示第j个综合指标在所有综合指标中所占的权重,Pj表示各种质第j个综合指标的方差贡献率。
2 结果与分析
2.1 干旱胁迫对各指标的影响
由图1可知,PEG-6000模拟干旱胁迫条件下,336份甜菜种质资源的胚轴直径、叶干重、株高、根长、叶鲜重、根鲜重、根干重、叶片饱和鲜重和叶片相对含水量等表型性状均极显著低于对照,干旱胁迫后根冠比、SS、SP和Pro含量等4个指标均极显著高于对照;SOD活性和MDA含量在干旱和对照处理下差异并不显著。不同甜菜种质的各指标在干旱胁迫后变化趋势不同,很难根据单一指标的变化情况客观、准确地比较不同甜菜种质耐旱情况,应该运用多指标综合评价的方法。
图1
图1
干旱胁迫对336份甜菜种质资源各指标的影响
Fig.1
Effects of drought stress on indexes of 336 sugar beet germplasm resources
2.2 主成分分析
由表1可知,前6个主成分因子可代表全部信息的75.95%,其中λ>0.918。根据结果可将原有的15个指标转化为6个相互独立的综合指标。第1主成分的贡献率为36.47%,其中胚轴直径、叶鲜重、根鲜重、根干重、叶干重和叶片饱和鲜重等指标的载荷系数绝对值较大,说明第1主成分主要反映的是生物量信息;第2主成分的贡献率为11.05%,其中SS和SP的载荷系数绝对值较大,说明第2主成分主要反映的是渗透调节物质的信息;第3主成分的贡献率为9.36%,其中根长、叶片相对含水量和根冠比的载荷系数较大,说明第3主成分主要反映的是干旱胁迫下会增长的表型指标;第4主成分的贡献率为6.68%,其在SOD上有较高载荷系数;第5主成分的贡献率为6.26%,其在MDA含量上有较高载荷系数;第6主成分的贡献率为6.12%,其在Pro含量上有较高载荷系数。
表1 甜菜种质资源各指标的主成分分析
Table 1
指标Index | F1 | F2 | F3 | F4 | F5 | F6 |
---|---|---|---|---|---|---|
胚轴直径EAD | 0.36 | 0.08 | 0.14 | 0.09 | 0.01 | 0.10 |
株高PH | 0.29 | -0.03 | 0.09 | -0.21 | 0.13 | -0.16 |
根长RL | 0.18 | -0.10 | 0.19 | -0.20 | -0.06 | -0.61 |
叶鲜重LFW | 0.40 | 0.04 | -0.14 | 0.00 | -0.03 | 0.03 |
根鲜重RFW | 0.38 | -0.07 | 0.21 | 0.11 | 0.02 | 0.13 |
根干重RDW | 0.34 | -0.02 | 0.18 | 0.19 | -0.00 | 0.20 |
叶干重LDW | 0.37 | 0.07 | -0.04 | 0.07 | -0.02 | -0.01 |
叶片饱和鲜重LSFW | 0.40 | 0.01 | -0.10 | 0.00 | -0.04 | 0.02 |
叶片相对含水量RLW | 0.14 | 0.17 | -0.47 | -0.16 | 0.12 | 0.15 |
根冠比RSR | -0.11 | -0.20 | 0.56 | 0.38 | 0.02 | 0.23 |
可溶性糖含量SS content | -0.02 | 0.55 | -0.16 | 0.31 | -0.36 | -0.10 |
可溶性蛋白含量SP content | -0.05 | 0.58 | 0.20 | 0.13 | -0.19 | 0.14 |
超氧化物歧化酶活性SOD activity | -0.01 | 0.23 | 0.30 | -0.72 | -0.02 | 0.46 |
脯氨酸含量Pro content | -0.01 | 0.34 | 0.38 | -0.16 | 0.02 | -0.47 |
丙二醛含量MDA content | -0.03 | 0.32 | 0.01 | 0.17 | 0.90 | -0.04 |
特征根Characteristic root | 5.47 | 1.66 | 1.41 | 1.00 | 0.94 | 0.92 |
累计贡献率Cumulative contribution rate (%) | 36.47 | 47.52 | 56.88 | 63.56 | 69.82 | 75.95 |
因子权重Factor weight | 0.48 | 0.15 | 0.12 | 0.09 | 0.08 | 0.08 |
由主成分分析(图2)可将336份种质大致分成3个类群,类群1是耐旱表现较好的种质,包括10、79、84、85、124、182、230、251、293和298号等10份;类群3是耐旱表现较差的种质,包括42、133、144、229和267号等5份;类群2的种质耐旱性介于类群1和类群3之间。
图2
图2
336份甜菜种质资源各指标主成分分析
Fig.2
Principal component analysis of indexes of 336 sugar beet germplasm resources
2.3 耐旱性综合评价
通过公式(1)计算耐旱系数,利用公式(2)和(3)计算各种质的综合指标值,再结合公式(4)计算各综合指标的权重,最后根据公式(5)得到各甜菜种质资源的耐旱性度量值(D)(图3),336份甜菜种质资源D值介于0.135~1.289,平均值为0.392,D值越大表示该甜菜种质耐旱性越强。耐旱性突出的种质有10、85和182号等,耐旱性弱的种质有42、133和310号。D值计算耐旱性的结果与主成分分析结果基本吻合。
图3
图3
336份甜菜种质资源D值散点图
Fig.3
Scatter plot of D values of 336 sugar beet germplasm resources
2.4 甜菜种质资源耐旱性聚类分析
对336份甜菜种质资源D值进行聚类分析(图4),共分为5类。第Ⅰ类为耐旱性强种质,有16份,占总数的4.76%,D值平均值为0.806;第Ⅱ类为耐旱性较强的种质,有49份,占总数的14.58%,D值平均值为0.564;第Ⅲ类为耐旱性中等的种质,有109份,占总数的32.44%,D值变化范围为0.363~0.501;第Ⅳ类为耐旱性较弱的种质,有79份,占总数的23.51%,D值平均值为0.326;第Ⅴ类为耐旱性弱的种质,有83份,占总数的24.70%,D值变化范围为0.135~0.283。
图4
图4
336份甜菜种质资源耐旱性分析聚类图
Fig.4
Cluster diagram of drought tolerance analysis of 336 sugar beet germplasm resources
2.5 D值与各指标的相关性分析
根据相关性分析结果(图5)可知,表型指标除叶片相对含水量和根冠比外其余各指标间均表现出显著正相关;根冠比与株高、叶鲜重、叶干重和叶片相对含水量呈显著负相关。表型指标与生理指标间除叶片相对含水量与SS间呈极显著相关外其余各指标间均无显著相关性。生理生化指标中渗透调节物质含量间呈极显著正相关,MDA含量与SP含量呈显著正相关,SOD活性与SP和Pro含量均呈显著正相关。D值与10个表型指标均表现出显著相关性,其中,D值与胚轴直径、株高、根长、叶鲜重、根鲜重、根干重、叶干重、叶片饱和鲜重和叶片相对含水量呈显著正相关,D值与根冠比呈显著负相关,D值与5个生理生化指标均无显著相关性。
图5
图5
耐旱性度量D值与各耐旱系数指标的相关性分析
Fig.5
Correlation analysis between D value of drought tolerance measure and drought tolerance coefficient of each index
3 讨论
3.1 耐旱性评价方法的选择
本研究通过以上方法得出各甜菜种质资源的D值与主成分分析结果高度符合,也从侧面印证了综合评价方法的可靠性。最终将甜菜种质资源的耐旱性分别为5个类群,筛选出耐旱型甜菜种质16份,这一结果与汪灿等[12]在薏苡中的研究结果一致。本研究鉴定出的耐旱材料可作为优良的甜菜种质资源加入到国家甜菜种质中期库的核心种质中,以便在今后新品种培育中加以利用。
3.2 干旱对甜菜种质资源表型和生理生化指标的影响
干旱胁迫可对植物的基因表达、代谢和生长变化产生影响,这些影响往往在几秒钟、几小时和几天内显现出来,但是这些变化中最先被观察到的是生长变化的影响[25],所以在干旱研究中表型指标至关重要。本研究的10个表型指标中有9个在经过干旱胁迫后是显著下降的,只有根冠比在干旱胁迫后显著增加,这表明干旱胁迫对表型指标的影响是显著的,可利用以上表型指标对甜菜种质资源进行耐旱性评价。
3.3 甜菜耐旱性评价指标的选择
植物的耐旱性是一个复杂的过程,不同的指标对干旱胁迫反应的敏感性和速度不同,植物在应对干旱胁迫时会随着胁迫的时间呈现出阶段性的特点,根系在感受到胁迫信号后,通过信号传导调节体内酶活性和渗透调节物质的表达,进而影响表型指标,使植物来适应外界胁迫[30⇓-32],本研究通过对336份甜菜种质资源胁迫7d后的各指标的耐旱系数和D值进行相关性分析,发现D值与所测的5个生理生化指标(SS、SP、SOD、Pro和MDA)均无显著相关性,与10个表型指标(EAD、PH、RL、LFW、RFW、RDW、LDW、LSFW、RLW和RSR)均表现出显著相关性,在长时间的干旱胁迫下甜菜种质耐旱鉴定可依据表型指标进行,这一结果与路之娟等[33]在苦荞中的研究结果相似。D值与根鲜重、胚轴直径和叶鲜重的相关系数分别为0.91、0.89和0.89,在大田试验中可以依据地上部指标叶鲜重、地下部指标根鲜重和胚轴直径这3个指标对甜菜种质耐旱性快速、有效地进行鉴定。
4 结论
本研究通过对336份甜菜种质资源苗期耐旱性进行分析,对D值进行聚类分析,筛选出16份苗期耐旱性强的甜菜种质。并根据相关性分析确定了D值与胚轴直径、株高、根长、叶鲜重、根鲜重、根干重、叶干重、叶片饱和鲜重和叶片相对含水量呈显著正相关,D值与根冠比呈显著负相关,本研究结果可以为甜菜抗旱育种提供材料和理论基础。
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Evaluation of some morpho- physiological characteristics in strawberry under different moisture stress regimes
,DOI:10.3233/JBR-180306 [本文引用: 1]
BACKGROUND: The strawberry is one of the most important small fruits; it is widely cultivated in the world and is sometimes subjected to drought stress. Investigating the effects of drought stress on cultivar related plant physiology is necessary to broaden the knowledge of resistance mechanisms to drought. OBJECTIVE: The present study was conducted to investigate the morpho-physiological characteristics of drought tolerance and the relationships between these and strawberry fruit yield under short- and long-term drought stress conditions. METHODS: The effect of two strawberry cultivars, Queen Eliza and Parus, was examined under four moisture regimes. A short-term stress for one month and a long-term stress for four months were undertaken in greenhouse condition. RESULTS: The analysis of variance indicated significant differences between the cultivars for carotenoid content, relative water content, relative water loss, stomatal conductance and quantum yield of photosystem II under short-term stress and stomatal conductance, carotenoid content and membrane stability under long-term stress. Correlation analysis revealed that yield correlated positively with relative water content and quantum yield of photosystem II ( Fv/Fm), under short-term and long-term stress. Using factor analysis, totally four factors were identified that accounted for 82.14% of the total variance. The factors were in decreasing order of influence yield components, photosynthesis, physiological characteristics and chlorophyll. CONCLUSIONS: Generally, 'Parus' was superior to 'Queen Eliza' in terms of measured traits under short and long-term stress conditions. It seems that measuring physiological traits is a convenient, simple and fast tool for screening genotypes for their drought tolerance.
Superoxide dismutase (SOD) activity of ceriops zippeliana in Segara Anakan Cilacap (Indonesia) under heavy metal accumulation
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Impact of foliar fertilization on the content of silicon and macronutrients in sugar beet
,DOI:10.3390/plants8050136 URL [本文引用: 1]
Turgor maintenance,osmotic adjustment and soluble sugar and proline accumulation in 49 pea cultivars in response to water stress
,DOI:10.1016/S0378-4290(98)00125-7 URL [本文引用: 3]
Inter-tissue and inter-organ signaling in drought stress response and phenotyping of drought tolerance
,DOI:10.1111/tpj.15619 URL [本文引用: 1]
Drought stress tolerance in seedlings of four deciduous species,common in nurseries of semi-arid region of Iran
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How reactive oxygen species and proline face stress together
,DOI:10.1016/j.plaphy.2014.04.007 PMID:24813727 [本文引用: 1]
Reactive oxygen species (ROS) are continuously generated as a consequence of plant metabolic processes due to incomplete reduction of O2. Previously considered to be only toxic by-products of metabolism, ROS are now known to act as second messengers in intracellular signalling cascades to trigger tolerance of various abiotic and biotic stresses. The accumulation of proline is frequently observed during the exposure of plants to adverse environmental conditions. Interestingly proline metabolism may also contribute to ROS formation in mitochondria, which play notably a role in hypersensitive response in plants, life-span extension in worms and tumor suppression in animals. Here we review current knowledge about the regulation of proline metabolism in response to environmental constraints and highlight the key role of ROS in the regulation of this metabolism. The impact of proline on ROS generation is also investigated. Deciphering and integrating these relationships at the whole plant level will bring new perspectives on how plants adapt to environmental stresses. Copyright © 2014 Elsevier Masson SAS. All rights reserved.
Genome-wide identification,structure characterization,expression pattern profiling,and substrate specificity of the metal tolerance protein family in Canavalia rosea (Sw.) DC
,DOI:10.3390/plants10071340 URL [本文引用: 1]
Osmoregulation and its actions during the drought stress in plants
,DOI:10.1111/ppl.13297 URL [本文引用: 1]
Wheat breeding highlights drought tolerance while ignores the advantages of drought avoidance:A meta-analysis
,DOI:10.1016/j.eja.2020.126196 URL [本文引用: 1]
Response mechanism of plants to drought stress
,DOI:10.3390/horticulturae7030050 URL [本文引用: 1]
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