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薏苡种质资源苗期抗旱性鉴定及抗旱指标筛选

本站小编 Free考研考试/2021-12-26

汪灿, 周棱波, 张国兵, 张立异, 徐燕, 高旭, 姜讷, 邵明波. 薏苡种质资源苗期抗旱性鉴定及抗旱指标筛选[J]. , 2017, 50(15): 2872-2887 https://doi.org/10.3864/j.issn.0578-1752.2017.15.003
WANG Can, ZHOU LingBo, ZHANG GuoBing, ZHANG LiYi, XU Yan, GAO Xu, JIANG Ne, SHAO MingBo. Drought Resistance Identification and Drought Resistance Indices Screening of Job’s Tears (Coxi lacryma-jobi L.) Germplasm Resources at Seedling Stage[J]. Scientia Acricultura Sinica, 2017, 50(15): 2872-2887 https://doi.org/10.3864/j.issn.0578-1752.2017.15.003

0 引言

【研究意义】薏苡(Coxi lacryma-jobi L.)是禾本科(Gramineae)薏苡属(Coix)一年生草本植物,具有丰富的营养价值和医药价值[1-2]。薏苡主要食用和药用部位为种仁,是一种高蛋白质、中脂肪、中糖的绿色食品,其营养堪称“禾本科植物之王”,具有健脾利湿、除脾止泻、清热解毒等功效[3]。干旱是影响薏苡生产的重要非生物胁迫因素[4]。因此,进行薏苡种质资源苗期抗旱性鉴定及抗旱指标筛选,对薏苡抗旱育种、抗旱机理及干旱调控缓解机制的研究具有重要意义。【前人研究进展】作物抗旱性鉴定及抗旱指标筛选是开展作物抗旱性研究首先要解决的关键技术,需要将不同指标相结合,对各个时期进行综合评价[5-7],其中,苗期鉴定因其时间短、容量大、重复性强、易于活体测定、环境影响小等优点[8],被广大研究者广泛采用。多年来,国内外****在作物苗期抗旱性鉴定及抗旱指标筛选方面开展了大量研究工作[9-11]。武斌等[12]结合SSR分子标记、简单相关分析和重心聚类方法研究53份玉米自交系的苗期耐旱性,认为叶片相对含水量和保水力是玉米苗期耐旱性的重要指标。邹成林等[13]通过相关分析和五级评分法综合评价玉米苗期抗旱性,认为相对含水量、叶绿素含量、硝酸还原酶活性、可溶性糖和丙二醛含量可作为玉米苗期的抗旱性鉴定指标。李真等[14]分析比较油菜DH群体各性状表现及其抗旱系数,并以抗旱系数对DH群体进行苗期抗旱性评价,认为地上部干重、根干重和总干重可作为甘蓝型油菜苗期抗旱性的主要评价指标。袁哲明等[15]基于支持向量回归非线性,筛选出了苗高、脯氨酸、丙二醛、叶龄、心叶下倒一叶面积和抗坏血酸6个水稻苗期抗旱性综合指标。王兰芬等[16]采用平均隶属函数法对70份绿豆种质进行苗期抗旱性综合评价,认为第1次旱胁迫幼苗存活率、萎蔫指数和株高为绿豆苗期抗旱性评价的适宜指标。李龙等[17]采用加权抗旱指数、抗旱度量值、相关分析、频次分析、灰色关联度分析、隶属函数分析和聚类分析相结合的方法,对50个普通菜豆品种进行苗期抗旱性鉴定,认为叶片相对含水量、PSⅡ最大量子产量和叶绿素含量可用于普通菜豆苗期抗旱性综合评价。【本研究切入点】在中国的大多数薏苡产区,虽然降水量充沛,但由于雨量不均,土壤保水能力较差,对薏苡生产不同程度上发生春旱和伏旱的威胁,干旱已成为制约薏苡生产的主要限制条件,目前,关于薏苡种质资源苗期抗旱性鉴定及抗旱指标筛选的研究鲜见报道。【拟解决的关键问题】本研究采用反复干旱胁迫法,在旱棚内进行盆栽试验,利用综合评价法对50份薏苡种质的幼苗干旱存活率、株高、茎粗、叶长、叶宽、地上部鲜重、地上部干重、根长、根粗、根鲜重和根干重进行鉴定与评价,从而筛选出苗期抗旱性强的薏苡种质及易测定的与薏苡种质抗旱性密切相关的指标,以期为薏苡抗旱育种、抗旱机理及干旱调控缓解机制的研究提供基础材料。

1 材料与方法

1.1 试验材料

供试薏苡种质共计50份,其中野生种3份、国审品种2份、黔薏苡1号选系3份、黔薏苡2号选系3份、云南地方品种3份、广西地方品种5份、四川地方品种3份、盘县地方品种10份、晴隆地方品种5份、兴仁地方品种5份、正安地方品种4份、安龙地方品种4份(电子版附表1)。

1.2 试验方法

试验于2016年4月至2016年7月在贵州省旱粮研究所旱棚内进行盆栽试验。在盆内(盆高50 cm、内径50 cm)装入20 cm厚的中等肥力水平的耕层土壤,土壤含有机质27.93 g·kg-1、全氮1.45 g·kg-1、全磷1.01 g·kg-1、全钾14.23 g·kg-1、碱解氮93.1 mg·kg-1、有效磷31.33 mg·kg-1、速效钾656.67 mg·kg-1、pH 7.6。播种前各盆施总养分≥45%的高效复合肥(含N 14%、P2O5 16%、K2O 15%)10 g,并浇水至田间持水量的80%(16.5%绝对含水量)。从每份薏苡种质中选取50粒大小均匀一致、饱满的种子,用0.1%的HgCl2消毒8 min后用蒸馏水冲洗5次,用滤纸吸干后用蒸馏水浸种24 h,然后按每盆50粒进行播种,播种后覆土2 cm。试验采用随机区组设计,设置正常供水(CK)和反复干旱(T)2个处理,3次重复。
第1次干旱-复水处理:出苗后正常供水,待幼苗长至四叶期时停止供水,当土壤绝对含水量降至田间持水量的20%(3.8%绝对含水量)时,复水至田间持水量的80%(16.5%绝对含水量)。
第2次干旱-复水处理:第1次复水后不再供水,当土壤绝对含水量降至田间持水量的20%(3.8%绝对含水量)时,复水至田间持水量的80%(16.5%绝对含水量)。

1.3 测定项目与方法

参照王兰芬等[16]方法,分别于第1次和第2次复水72 h后调查存活率,以幼苗叶片转为鲜绿色为存活。幼苗干旱存活率(seedling drought survival rate,DS)=(XDS1/XTT×100%+XDS1/XTT×100%)/2。式中XDS1XDS2分别为第1次和第2次复水后存活苗数,XTT为第1次干旱前总苗数。
于第2次复水72 h后,每盆随机选择5株植株,洗净后将根系剪下与幼苗分开。量取主茎自地面至植株顶端的距离,即为株高(plant height,PH);主茎中部最长节间中部的直径(不包括叶鞘),即为茎粗(culm diameter,CD);主茎中部最大叶片基部至叶尖的长度,即为叶长(leaf length,LL);主茎中部最大叶片最宽处的长度,即为叶宽(leaf width,LW);主根的长度,即为根长(root length,RL);第1侧根发根处主根的直径,即为根粗(root diameter,RD);分别称取幼苗和根系鲜物质重量,即为地上部鲜重(shoot fresh weight,SFW)和根鲜重(root fresh weight,RFW);将幼苗和根系晾干,于60℃下烘干后分别称重,即为地上部干重(shoot dry weight,SDW)和根干重(root dry weight,RDW)。

1.4 数据统计分析

用Microsoft Excel 2013整理数据,用SPSS 19进行统计分析。参照谢小玉等[18]的方法,采用配对处理t检验对各指标测定值进行平均数差异显著性检测。按公式(1)和(2)分别计算单项抗旱系数(drought resistance coefficient,DC)和综合抗旱系数(comprehensive drought resistance coefficient,CDC)。式中xiCKi分别表示反复干旱和正常供水处理的指标测定值。
$$DC=\frac{x_i}{CK_i} \ \ (1)$$
$$CDC=\frac{1}{n}\sum^n_{i=1}DC\ \ (2)$$
参照罗俊杰等[19]和兰巨生[20]方法,针对各指标DC值,进行简单相关分析、连续变数次数分布统计分析和主成分分析。按公式(3)、(4)和(5)分别计算因子权重系数(ωi)、各基因型各综合指标的隶属函数值[μ(xi)]和抗旱性度量值(drought resistance comprehensive evaluation value,D)。式中Pi为第i个综合指标贡献率,表示第i个指标在所有指标中的重要程度,xiximaxximin分别表示第i个综合指标及第i个综合指标的最大值和最小值。
$$\omega _i=P_i \div \sum^n_{i=1}P_i\ \ (3)$$
$$\mu(x_i)=\frac{x_i-x_{imin}}{x_{imax}-x_{imin}}\ \ (4)$$
$$D=\sum^n_{i=1}[\mu(x_i)\times(P_i\div \sum^n_{i=1}P_i)]\ \ (5)$$
参照孟庆立等[21]和祁旭升等[22]方法,以各指标DC值为比较序列,D值为参考序列进行灰色关联度分析,获得各指标DC值与D值间的关联度(γD),按公式(6)和(7)分别计算各指标权重系数[ωi(γ)]和加权抗旱系数(weight drought resistance coefficient,WDC)。式中γi为各指标关联度。
$$\omega_i(\gamma)=\gamma _i\div \sum^n_{i=1}\gamma _i \ \ (6)$$
$$WDC=\sum^n_{i=1}[DC\times(\gamma _i\div \sum^n_{i=1}\gamma _i)]\ \ (7)$$
参照朱宗河等[23]和张彦军等[24]方法,以各指标DC值为比较序列,WDC值为参考序列进行灰色关联度分析,获得各指标DC值与WDC值间的关联度(γWDC)。最后针对供试薏苡种质D值,采用欧式距离和加权配对算术平均法(weighted pair group method average,WPGMA)进行聚类分析,划分抗旱级别,并分别以D值、CDC值和WDC值为参考序列,对各指标DC值进行逐步回归分析,求取回归方程。

2 结果

2.1 供试种质的代表性及其指标测定值分析

干旱胁迫对供试种质各指标测定值均有显著影响,处理间和种质间的差异均达显著水平(表1)。种质间变异系数为0.134—0.747,说明本试验所选薏苡种质类型丰富,所选指标对干旱胁迫反应较敏感,干旱胁迫处理效果好,具有较好的代表性。此外,供试种质各指标在反复干旱和正常供水处理下的测定值相关系数介于0.621—0.945,这进一步说明各指标对干旱胁迫反应的敏感性存在差异,采用各指标测定值难以直接考量其抗旱性。
Table 1
表1
表1反复干旱和正常供水条件下供试薏苡种质各指标测定值及其均值差异性分析
Table 1Measured values of all indices in tested Job’s tears germplasms under repeated drought and normal water supply and its mean variance analysis
编号
Number
幼苗干旱存活率
DS (%)
株高
PH (cm)
茎粗
CD (mm)
叶长
LL (cm)
叶宽
LW (cm)
地上部鲜重
SFW (g)
地上部干重
SDW (g)
根长
RL (cm)
根粗
RD (mm)
根鲜重
RFW (g)
根干重
RDW (g)
CKTCKTCKTCKTCKTCKTCKTCKTCKTCKTCKT
YG0193.3721.7219.136.315.479.628.900.990.872.662.400.240.191.931.774.703.820.500.410.120.09
YG0290.8326.0322.255.364.7017.0313.301.771.344.803.950.600.443.122.626.425.280.520.340.120.07
YG0386.8024.2819.706.574.6915.2310.601.690.844.133.330.520.372.732.385.763.810.400.290.090.06
YG0434.2019.5815.607.044.5110.888.731.000.441.180.740.150.081.050.626.183.670.100.050.020.01
YG0593.0322.4515.304.692.8313.7510.201.030.504.071.230.510.141.971.034.112.300.200.070.050.02
YG0630.5622.2013.634.581.9913.556.071.100.222.290.930.290.101.550.724.011.610.310.240.070.05
YG0754.2224.1018.824.872.379.936.230.980.431.901.200.240.132.080.683.211.170.230.140.050.03
YG0851.3025.3319.354.632.8210.207.350.900.312.171.340.270.151.621.134.062.290.170.080.040.02
YG0986.7323.3015.804.132.1410.456.300.720.471.210.890.150.101.120.823.621.740.220.120.050.03
YG1084.8323.5715.132.581.7220.2017.321.030.652.040.460.450.121.870.732.261.390.110.050.020.01
YG1160.5623.7712.702.001.389.337.431.100.501.621.030.200.112.351.881.751.120.100.030.020.01
YG1283.7727.4217.253.271.7814.928.471.090.832.331.530.290.171.850.622.871.450.160.080.040.02
YG1363.8922.3017.303.662.768.786.631.140.481.491.270.190.142.400.913.212.240.080.020.020.01
YG1447.8020.3214.282.791.715.723.650.820.561.320.780.170.092.171.302.451.390.290.150.070.03
YG1546.3726.5019.933.172.078.085.181.030.652.411.250.300.142.921.072.781.680.160.060.040.01
YG1677.4325.4518.703.462.309.084.781.100.362.231.850.280.211.600.803.041.870.210.100.050.02
YG1780.4323.3316.583.902.489.336.551.120.851.891.320.240.151.030.633.422.020.130.030.060.02
YG1882.1724.8317.454.563.798.005.872.641.861.811.350.230.151.571.074.003.080.240.140.050.03
YG1974.1723.0313.354.291.437.875.151.050.391.210.920.150.101.901.424.271.620.060.020.030.01
YG2048.7223.9517.656.133.646.254.271.050.171.791.390.220.151.501.205.372.960.160.080.040.02
YG2150.7224.6515.386.023.307.936.531.050.312.051.010.260.112.231.385.282.680.140.070.030.02
YG2245.8326.1715.954.882.419.407.671.200.352.482.060.310.231.420.804.281.960.280.110.060.02
YG2388.3722.0016.583.371.569.326.251.581.182.411.770.300.202.371.452.951.270.230.170.050.04
YG2461.4422.6719.403.882.678.134.951.230.771.781.530.220.171.701.253.402.170.170.100.040.02
YG2560.3326.3516.773.971.857.554.130.980.281.881.050.240.122.331.352.360.710.210.110.050.02
YG2690.3721.0515.223.281.278.906.250.970.481.880.820.240.092.271.352.881.030.130.070.030.02
YG2793.7725.2822.624.574.228.137.650.900.813.072.920.380.323.573.354.003.440.160.140.040.03
YG2881.6027.8315.584.743.977.805.150.970.352.652.020.330.221.720.674.163.230.150.090.030.02
YG2984.6721.6515.884.133.228.676.281.130.801.561.120.200.122.151.223.622.620.200.120.050.03
YG3077.6323.1815.283.221.769.587.451.280.601.551.270.190.141.400.402.821.430.170.070.040.02
YG3178.8322.8719.723.611.999.275.831.000.821.441.060.180.121.170.703.161.610.200.120.050.03
YG3281.4125.6319.272.551.879.226.821.050.602.031.430.250.161.030.702.241.130.150.100.030.02
YG3346.0024.7020.955.153.587.555.380.970.693.242.280.400.251.420.804.512.910.200.160.050.03
YG3448.8727.1723.874.133.159.286.421.271.121.951.670.240.192.071.533.622.560.170.090.040.02
YG3585.1023.7016.004.563.5510.806.731.150.651.951.720.660.201.651.474.002.890.110.040.030.01
YG3691.9324.0211.906.365.428.906.731.070.642.482.060.310.232.051.605.584.400.260.170.060.04
YG3791.8022.1518.577.326.4910.089.331.050.905.304.940.330.271.851.706.695.580.180.160.040.04
YG3881.6029.3825.252.491.4310.055.851.170.351.591.270.200.141.751.072.181.160.260.090.060.02
YG3946.7824.2017.303.532.879.726.301.080.242.412.050.300.233.032.173.102.330.190.120.040.03
YG4051.0225.3217.526.654.439.487.821.750.514.303.720.540.413.872.505.833.600.350.230.080.05
YG4190.5024.5313.274.013.318.124.801.230.731.401.110.180.122.070.883.522.690.270.100.050.02
YG4276.0625.8318.232.641.8212.209.581.700.942.602.000.330.223.152.402.321.090.150.100.030.02
YG4391.8323.4312.652.912.1610.078.251.660.441.531.350.190.152.771.672.551.760.170.110.040.02
YG4432.5831.4824.203.312.0814.9011.321.250.854.743.640.590.404.222.972.901.690.330.160.080.04
YG4588.4030.9526.777.636.5714.3010.241.731.195.233.540.650.394.123.055.274.040.480.320.110.07
YG4634.2324.6513.323.661.448.324.351.200.183.591.050.260.051.420.373.761.160.180.040.030.01
YG4787.4030.5021.986.805.2718.009.471.471.044.513.460.560.383.352.535.964.280.310.220.070.05
YG4878.6035.7023.136.315.4716.4514.801.911.495.763.110.720.353.922.335.534.450.700.480.160.10
YG4959.8734.4725.805.493.8414.7711.801.530.854.733.750.590.423.481.454.823.120.390.200.090.04
YG5029.6030.8313.842.690.8813.374.951.110.121.660.330.210.042.430.522.830.780.120.020.040.01
平均值
Average
69.5712.1217.844.433.0110.577.441.220.662.571.800.320.192.211.383.872.400.230.130.050.03
变异系数CV0.2920.1340.2050.3230.4760.3000.3730.2810.5410.4820.5810.4820.5420.3800.5390.3250.4980.5510.7470.5290.712
标准误
SE
0.4340.0950.2380.0390.0870.0130.0650.0810.0060.002
t16.7815.0913.1614.508.759.8112.6418.0914.9013.18
P0.0001**0.0001**0.0001**0.0001**0.0001**0.0001**0.0001**0.0001**0.0001**0.0001**
相关系数R0.6210.8910.8490.6960.8680.8120.8360.8920.9450.923

CK: Normal water supply treatments; T: Repeated drought treatments. ** is significant difference at 0.01 probability levelCK:正常供水处理;T:反复干旱处理。**表示0.01水平差异显著
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2.2 单项指标分析

与正常供水处理相比,供试种质在反复干旱处理后,各指标均发生不同程度变化(表2)。同一指标各种质的DC值差异明显,变异系数介于0.156—0.405,但不同种质间DC值所反映的抗旱性不同,且同一种质各指标的DC值存在较大差异,说明各指标对干旱胁迫反应的敏感性各异。
相关分析表明(表3),各指标都至少与一个其它指标呈显著或极显著相关,说明各指标间存在一定程度的相关性。其中,幼苗干旱存活率与茎粗、叶宽和根粗呈极显著正相关,与叶长、地上部鲜重、地上部干重、根长、根鲜重和根干重呈显著正相关,与株高呈不显著的正相关。
此外,同一区间各指标DC值分布次数和频率相差较大(表4)。DC>0.6的幼苗干旱存活率、株高、茎粗、叶长、叶宽、地上部鲜重、地上部干重、根长、根粗、根鲜重和根干重的分布频率分别为66%、84%、68%、82%、42%、78%、66%、56%、52%、38%和32%,说明各指标对干旱胁迫反应的敏感性依次为根干重、根鲜重、叶宽、根粗、根长、幼苗干旱存活率、地上部干重、茎粗、地上部鲜重、叶长和株高。因此,直接采用这些指标会由于指标间信息的重叠,很难客观、准确、有效地评价各种质的抗旱性,从而影响抗旱鉴定结果。
Table 2
表2
表2供试薏苡种质各指标的抗旱系数
Table 2Drought resistance coefficients of all indices in tested Job’s tears germplasms
编号
Number
幼苗干旱存活率
DS
株高
PH
茎粗
CD
叶长
LL
叶宽
LW
地上部鲜重
SFW
地上部干重
SDW
根长
RL
根粗
RD
根鲜重
RFW
根干重
RDW
YG010.9340.8810.8680.9250.8820.9020.7850.9140.8130.8070.772
YG020.9080.8550.8780.7810.7570.8240.7320.8410.8220.6500.622
YG030.8680.8110.7140.6960.5010.8070.7170.8710.6620.7180.687
YG040.3420.7970.6410.8020.4370.6290.5590.5870.5940.4360.417
YG050.9300.6820.6030.7420.4870.3030.2690.5250.5590.3620.346
YG060.3060.6140.4340.4480.1970.4060.3610.4620.4020.7900.755
YG070.5420.7810.4860.6280.4340.6290.5590.3280.3630.6120.585
YG080.5130.7640.6090.7210.3480.6190.5500.7010.5650.4580.438
YG090.8670.6780.5180.6030.6600.7340.6520.7310.4800.5420.518
YG100.8480.6420.6660.8570.6340.2240.2590.3930.6170.4890.468
YG110.6060.5340.6890.7960.4580.6340.5640.8010.6390.3040.291
YG120.8380.6290.5440.5680.7590.6550.5820.3330.5050.5160.493
YG130.6390.7760.7540.7550.4170.8560.7600.3790.6990.2490.520
YG140.4780.7030.6130.6380.6820.5920.5260.5980.5680.5080.486
YG150.4640.7520.6540.6410.6320.5180.4600.3660.6060.3990.382
YG160.7740.7350.6650.5270.3280.8300.7380.5020.6160.4610.441
YG170.8040.7110.6370.7020.7610.6960.6180.6130.5900.2370.374
YG180.8220.7030.8310.7330.7050.7470.6640.6810.7700.5900.564
YG190.7420.5800.3320.6550.3680.7590.6740.7460.3790.2800.382
YG200.4870.7370.5930.6830.1650.7730.6870.8020.5500.4730.452
YG210.5070.6240.5470.8240.2940.4940.4390.6190.5070.5030.481
YG220.4580.6100.4930.8160.2940.8300.7380.5650.4570.3910.343
YG230.8840.7540.4640.6710.7480.7330.6510.6130.4300.7230.691
YG240.6140.8560.6890.6090.6220.8570.7610.7350.6390.6030.573
YG250.6030.6360.4670.5470.2900.5570.4950.5790.3020.5100.487
YG260.9040.7230.3860.7020.4970.4350.3870.5960.3570.5740.549
YG270.9380.8950.9230.9410.9070.9500.8450.9390.8580.9170.877
YG280.8160.5600.8380.6600.3590.7610.6760.3880.7770.5760.551
YG290.8470.7340.7800.7250.7060.7160.6370.5660.7230.5900.564
YG300.7760.6590.5450.7770.4700.8210.7300.2860.5050.4470.428
YG310.7880.8620.5510.6290.8170.7400.6580.6030.5100.6200.593
YG320.8140.7520.7310.7400.5760.7040.6260.6770.5050.6810.651
YG330.4600.8480.6950.7130.7120.7030.6250.5650.6440.7860.752
YG340.4890.8790.7640.6910.8820.8600.7640.7420.7080.5070.485
YG350.8510.6750.7790.6230.5620.8830.3050.8890.7220.3520.337
YG360.9190.4950.8510.7570.5950.8320.7400.7800.7890.6660.637
YG370.9180.8380.8870.9260.8570.9330.8020.9190.8340.8700.832
YG380.8160.8590.5720.5820.2990.7970.7090.6100.5300.3410.326
YG390.4680.7150.8120.6480.2230.8500.7560.7140.7520.5970.570
YG400.5100.6920.6660.8240.2910.8650.7690.6470.6170.6700.641
YG410.9050.5410.8250.5910.5950.7900.7020.4270.7650.3590.364
YG420.7610.7060.6880.7860.5510.7680.6820.7620.4700.6940.664
YG430.9180.5400.7440.8200.2630.8850.7860.6020.6890.6410.613
YG440.3260.7690.6290.7600.6830.7670.6820.7040.5830.4910.469
YG450.8840.8650.8600.7160.6890.6760.6010.7400.7670.6640.635
YG460.3420.5400.3920.5230.1470.2910.1990.2590.3080.2010.226
YG470.8740.7210.7750.5260.7070.7680.6820.7560.7180.6960.665
YG480.7860.6480.8680.8980.7800.5410.4810.5960.8040.6810.651
YG490.5990.7490.6990.7990.5580.7920.7040.4160.6470.5110.489
YG500.2960.4490.3260.3700.1050.2010.1780.2120.2740.1530.192
平均值 Average0.6960.7110.6590.7020.5340.6990.6110.6140.6000.5380.527
变异系数 CV0.2920.1560.2340.1740.4050.2600.2700.2980.2540.3240.287


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Table 3
表3
表3供试薏苡种质各指标抗旱系数的相关性
Table 3Correlations of drought resistance coefficients of all indices in tested Job’s tears germplasms
指标
Indices
幼苗干旱存活率
DS
株高
PH
茎粗
CD
叶长
LL
叶宽
LW
地上部鲜重
SFW
地上部干重
SDW
根长
RL
根粗
RD
根鲜重
RFW
株高 PH0.130
茎粗 CD0.444**0.341*
叶长 LL0.297*0.283*0.526**
叶宽 LW0.500**0.531**0.521**0.394**
地上部鲜重 SFW0.328*0.415**0.550**0.334*0.313*
地上部干重 SDW0.288*0.426**0.500**0.377**0.307*0.911**
根长 RL0.344*0.432**0.479**0.421**0.385**0.558**0.454**
根粗 RD0.437**0.326*0.962**0.524**0.525**0.561**0.509**0.479**
根鲜重 RFW0.329*0.450**0.463**0.381**0.458**0.367**0.426**0.477**0.406**
根干重 RDW0.354*0.478**0.485**0.402**0.479**0.413**0.481**0.464**0.433**0.960**

* and ** are significant correlation at 0.05 and 0.01 probability level, respectively. The same as below*和**分别表示0.05和0.01水平显著相关。下同
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Table 4
表4
表4供试薏苡种质各指标抗旱系数在不同区间的分布
Table 4Different distributions of drought resistance coefficients of all indices in tested Job’s tears germplasms
指标
Indices
0<DC<0.20.2<DC<0.40.4<DC<0.60.6<DC<0.80.8<DC<1
次数
Times
频率
Freq. (%)
次数
Times
频率
Freq. (%)
次数
Times
频率
Freq. (%)
次数
Times
频率
Freq. (%)
次数
Times
频率
Freq. (%)
幼苗干旱存活率 DS00510122410202346
株高 PH000081631621122
茎粗 CD0048122423461122
叶长 LL001281631621020
叶宽 LW48112214281632510
地上部鲜重 SFW004871423461632
地上部干重 SDW245101020316224
根长 RL0091813262040816
根粗 RD0061218362142510
根鲜重 RFW1211221938163236
根干重 RDW1210202346142824

Freq.: Frequency
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2.3 主成分分析

各因子特征值中前6个因子的累计贡献率达91.40%,其特征根λ>0.613(表5)。因此,抽取前6个因子,将具有相同本质的变量归为一类,可将原来各单项指标转换成6个新的相互独立的综合指标(分别用F1、F2、F3、F4、F5和F6表示)。F1在茎粗上有较高载荷量,F2在根粗、根鲜重和根干重上有较高载荷量,F3在地上部鲜重和地上部干重上有较高载荷量,F4在株高和叶宽上有较高载荷量,F5在幼苗干旱存活率上有较高载荷量,F6在根长上有较高载荷量。
Table 5
表5
表5供试薏苡种质各指标主成分的特征向量及贡献率
Table 5Eigenvectors and contribution rates of principal components of all indices in tested Job’s tears germplasms
指标
Indices
因子载荷 Factor loading
F1F2F3F4F5F6
幼苗干旱存活率 DS0.235-0.1010.426-0.1860.6940.224
株高 PH0.2570.299-0.2390.688-0.026-0.064
茎粗 CD0.346-0.2900.247-0.034-0.230-0.343
叶长 LL0.266-0.0970.207-0.022-0.5420.533
叶宽 LW0.2900.1590.3440.4790.237-0.088
地上部鲜重 SFW0.316-0.321-0.467-0.0520.199-0.017
地上部干重 SDW0.312-0.233-0.493-0.1060.143-0.081
根长 RL0.298-0.005-0.1340.0530.0060.627
根粗 RD0.340-0.3450.2410.006-0.220-0.324
根鲜重 RFW0.3120.518-0.022-0.359-0.068-0.110
根干重 RDW0.3250.487-0.040-0.340-0.045-0.139
特征根 Characteristic root5.5731.2141.1290.7750.7500.613
贡献率 Contribution rate (%)50.6611.0310.267.056.825.57
累计贡献率 Cumulative contribution rate (%)50.6661.7071.9679.0185.8391.40
因子权重 Factor weight0.5540.1210.1120.0770.0750.061


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2.4 供试种质的综合抗旱性评价

供试种质CDC值和WDC值分别介于0.251—0.908和0.149—0.908,平均值分别为0.626和0.624,变异系数分别为0.185和0.186,根据CDC值和WDC值的大小对供试种质进行抗旱性排序,其结果基本相同(表6)。其中,抗旱性强的种质有yy18-1、yy14-3和yy13-1,抗旱性弱的种质有yy12-7,其余种质介于两者之间。
供试种质D值介于0.196—0.775,平均值为0.517,变异系数为0.206,根据D值的大小对供试种质进行抗旱性排序,其抗旱性强的种质有yy18-1、yy14-3和yy13-1,抗旱性弱的种质有yy12-7,其余种质介于两者之间(表6)。这与基于CDC值和WDC值的供试种质抗旱性强弱评价结果基本吻合。
Table 6
表6
表6供试薏苡种质抗旱性评价的CDC值、WDC值及D值
Table 6CDC value, WDC value, and D value of drought resistance evaluation in tested Job’s tears germplasms
编号
Number
隶属函数 Subordinate function valueCDC值
CDC value
排序
Rank
D值
D value
排序
Rank
WDC值
WDC value
排序
Rank
μ1μ2μ3μ4μ5μ6
YG010.9260.5330.4220.6230.4510.6250.86230.74520.8613
YG020.8140.3880.4340.6920.5580.4450.78840.66940.7864
YG030.7280.5520.2390.4650.6650.5700.73270.61760.7297
YG040.5050.4130.2440.8000.0000.5030.567380.450390.56338
YG050.4070.4440.8430.6780.5550.6970.528460.510240.52646
YG060.3511.0000.2110.1640.3370.1040.470480.383480.47348
YG070.4450.7500.1800.5670.5530.2550.541420.458340.53941
YG080.5000.4330.2470.6690.2670.6160.571370.466330.56737
YG090.5550.5110.3020.5431.0000.6430.635220.559190.63322
YG100.4550.5571.0000.5840.2680.5450.554400.530230.55639
YG110.4890.0840.4660.5300.2870.8350.574360.447420.56936
YG120.4810.5060.4820.5440.9880.1780.584330.508250.58431
YG130.5780.1370.2190.6760.3930.1690.618280.453380.61228
YG140.5020.5290.3730.7110.4220.3430.581350.491280.58033
YG150.4390.4410.4720.8560.3300.1170.534440.448400.53343
YG160.5320.2700.1430.5160.8410.1570.601300.456350.59630
YG170.5320.2230.4660.8530.7660.5690.613290.532220.60929
YG180.6940.3360.5040.5390.5390.3290.71090.584120.7099
YG190.4090.3010.1190.4810.9001.0000.536430.442430.52944
YG200.5190.3460.0000.5130.3290.6750.582340.435450.57535
YG210.4370.5050.4170.4270.0930.7690.531450.437440.52845
YG220.4540.2210.0480.4730.3070.7630.545410.390470.53742
YG230.6100.7840.2990.5100.9590.5810.669190.61370.66919
YG240.6660.4890.0900.7340.6300.2930.687130.560180.68413
YG250.3670.6250.2140.4140.7000.5700.498470.422460.49547
YG260.4390.7860.5200.5360.7920.8520.555390.549200.55440
YG271.0000.5920.3920.5380.4070.5450.90810.77510.9081
YG280.5880.2070.4640.1680.5280.0000.633240.455370.63123
YG290.6620.4040.5160.5780.5960.2730.690120.579140.68912
YG300.5030.2930.2700.4770.6640.4110.586310.456360.58132
YG310.6190.6830.2760.8030.8850.3980.670180.60980.66918
YG320.6430.6060.3690.4700.5900.5350.678170.584110.67717
YG330.6720.7770.2610.6390.2440.1320.682150.571160.68314
YG340.6970.3770.1831.0000.4390.2800.706100.580130.70411
YG350.5660.1430.5440.7100.7110.6440.634230.539210.63024
YG360.7220.2310.5100.0970.5990.4660.73360.566170.7316
YG370.9460.5490.4080.4920.4130.5800.87420.74130.8742
YG380.5040.2680.0890.7910.8780.5040.586320.479300.57834
YG390.6290.2870.0620.3720.2120.1990.646210.447410.64121
YG400.6340.4320.0700.2880.1620.5100.654200.477310.64920
YG410.5530.0000.5290.4330.8510.0560.624260.467320.62126
YG420.6530.5870.2740.3860.5390.7060.685140.577150.68215
YG430.6560.2220.3210.0000.5270.5350.682160.498260.67716
YG440.5790.4310.1450.8180.1950.5060.624270.497270.62027
YG450.7380.5200.5170.6880.5420.3220.73650.64350.7365
YG460.0980.4550.4430.5800.3560.3900.312490.254490.30949
YG470.6950.5140.3790.4670.8960.1720.71780.603100.7178
YG480.6910.4840.8230.4480.1550.3900.703110.60490.70510
YG490.5920.3360.2920.6260.3270.2760.633250.491290.62925
YG500.0000.4570.4340.4980.5180.2400.251500.196500.24950
平均值
Average
0.6260.5170.624
变异系数
CV
0.1850.2060.186

μ1, μ2, μ3, μ4, μ5, and μ6 are subordinate function values of six factors, respectivelyμ1μ2μ3μ4μ5μ6分别表示6个因子的隶属函数值
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2.5 灰色关联度分析

各指标DC值与D值间的关联度大小依次为根干重、叶宽、根鲜重、茎粗、根粗、幼苗干旱存活率、根长、株高、地上部鲜重、地上部干重、叶长(表7),反映了各指标DC值与D值的密切程度,这与各指标对干旱胁迫反应的敏感性基本吻合。
此外,各指标DC值与WDC值间的关联度大小依次为根干重、根鲜重、茎粗、根粗、叶宽、地上部鲜重、地上部干重、根长、株高、幼苗干旱存活率、叶长(表7),这与各指标DC值与D值的密切程度基本吻合。

2.6 聚类分析及抗旱级别的划分

λ=10处将50份供试种质分为5类(图1)。其中第Ⅰ类为高度抗旱型种质,有yy18-1、yy14-3和yy13-1共3份,占总数的6%;第Ⅱ类为抗旱型种质,共17份,占总数的34%;第Ⅲ类为中等抗旱型种质,共9份,占总数的18%;第Ⅳ类为敏感型种质,共20份,占总数的40%;第Ⅴ类为高度敏感型种质,有yy12-7共1份,占总数的2%。
Table 7
表7
表7供试薏苡种质各指标DC值与D值和WDC值的关联度及各指标权重
Table 7Correlation degree between DC value of all indices and D value together with WDC value and indices weight in tested Job’s tears germplasms
指标 Indices关联度 γD排序 Rank权重系数 Weight关联度 γWDC排序 Rank
幼苗干旱存活率 DS0.71560.0900.70710
株高 PH0.69780.0880.7149
茎粗 CD0.73840.0930.7743
叶长 LL0.681110.0860.68911
叶宽 LW0.76920.0970.7405
地上部鲜重 SFW0.68390.0860.7396
地上部干重 SDW0.682100.0860.7377
根长 RL0.70470.0890.7228
根粗 RD0.72550.0910.7604
根鲜重 RFW0.76430.0960.7782
根干重 RDW0.77010.0970.7841


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根据供试种质的抗旱性聚类分析及抗旱级别划分
显示原图|下载原图ZIP|生成PPT
图1基于D值的供试薏苡种质抗旱性系统聚类图
-->Fig. 1Fuzzy clustering dendrogram of drought resistance in tested Job’s tears germplasms based on D value
-->

结果,对供试种质抗旱性评价指标进行分级统计,结果表明(表8),除叶长、地上部鲜重、地上部干重和根干重外,其余指标的隶属函数值、CDC值、D值和WDC值均随抗旱级别的升高而增大。此外,CDC值、D值和WDC值在不同抗旱级别上的差异较大,可为其他薏苡种质抗旱级别的划分提供依据。
Table 8
表8
表8供试薏苡种质抗旱性评价指标的分级
Table 8Classification of drought resistance evaluation indices in tested Job’s tears germplasms
指标
Indices
隶属函数 Subordinate function value
IIIIIIIVV
幼苗干旱存活率 DS0.9880.7750.6800.4440.000
株高 PH0.9480.6940.5130.5080.000
茎粗 CD0.9480.6400.5540.4600.000
叶长 LL0.9820.5860.6190.5300.000
叶宽 LW0.9690.7130.6150.3080.000
地上部鲜重 SFW0.9710.7090.5910.6460.000
地上部干重 SDW0.9480.7090.5210.6420.000
根长 RL0.9790.6670.4830.4490.000
根粗 RD0.9610.6190.5740.4670.000
根鲜重 RFW0.9310.6450.3960.3920.000
根干重 RDW0.9270.6210.3800.3820.000
CDC值 CDC value0.8810.6930.6040.5600.251
D值 D value0.7530.5920.5110.4360.196
WDC值 WDC value0.8810.6920.6010.5560.249

Ⅰ, Ⅱ, Ⅲ, Ⅳ, and Ⅴ represent different drought resistance levelsⅠ、Ⅱ、Ⅲ、Ⅳ、Ⅴ表示不同抗旱级别
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2.7 抗旱指标的筛选

分别以D值、CDC值和WDC值为参考序列,对各指标DC值进行逐步回归分析,得到的3个回归方程的决定系数R2≈1,F检验均达极显著水平(表9)。说明模型拟合度好,回归方程最优,其解释能力强,预测精度高,用这3个方程进行薏苡种质资源苗期抗旱性评价效果好。
根据D值与各指标DC值的回归方程可知(表9),在薏苡种质资源苗期抗旱性鉴定中,有选择性地测定与D值密切相关的指标,如幼苗干旱存活率、株高、叶宽、根长和根鲜重,可有效鉴定薏苡种质资源的抗旱性,从而使鉴定工作简化。
Table 9
表9
表9供试薏苡种质抗旱性模型预测
Table 9Model predict of drought resistance in tested Job’s tears germplasms
因变量
Dependent variable
多元逐步回归方程
Multiple stepwise regression equation
决定系数
R2
FP统计量
Statistic
D值 D valuey=-0.001+0.17x1+0.20x2+0.18x5+0.13x8+0.16x100.995845.32**0.00012.46
CDC值 CDC valuey=0.05+0.08x1+0.18x3+0.08x4+0.12x5+0.20x7+0.12x8+0.15x100.996685.67**0.00012.32
WDC值 WDC valuey=0.05+0.08x1+0.19x3+0.08x4+0.12x5+0.19x7+0.12x8+0.17x100.996725.26**0.00012.32

x1: Seedling drought survival rate; x2: Plant height; x3: Culm diameter; x4: Leaf length; x5: Leaf width; x7: Shoot dry weight; x8: Root length; x10: Root fresh weightx1:幼苗干旱存活率;x2:株高;x3:茎粗;x4:叶长;x5:叶宽;x7:地上部干重;x8:根长;x10:根鲜重
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3 讨论

3.1 薏苡种质资源苗期抗旱性评价方法

作物抗旱性的综合评价不仅需要选择适宜的评价指标,而且要有合适的评价方法。一般认为,多指标多方法相结合的综合评价比较可靠[25-30]。对于作物抗旱性评价的方法,大多数采用等权重的评价方法,却忽视了各项指标的不同重要程度。本研究采用D值、CDC值和WDC值等综合评价指标,结合单项指标抗旱系数、相关分析、频次分析、主成分分析、灰色关联度分析、隶属函数分析、聚类分析及逐步回归分析,对薏苡种质资源的抗旱性综合评价值作出评判,消除因各指标单位不同带来的差异,同时结合指标变异系数来确定每一个指标在抗旱性评价体系中的权重,对与抗旱性相关密切的指标分配较高的比重。以D值为主要综合评价指标,以CDC值和WDC值作为辅助综合评价指标的评价方法,既考虑了各指标间的相互关系,又考虑到各指标的重要性,评价结果客观、可靠。

3.2 薏苡种质资源苗期抗旱性的鉴定

作物抗旱性鉴定的最终结果是要划分供试种质的抗旱等级,以此来判定其抗旱能力[22]。在本研究中,针对D值,将供试薏苡种质划分为高度抗旱型种、抗旱型种质、中等抗旱型种质、敏感型种质和高度抗旱型种质5类,这与罗俊杰等[19]在胡麻上的研究结果基本一致。此外,在本研究中,鉴定出抗旱性强的种质有yy18-1、yy14-3和yy13-1,抗旱性弱的种质有yy12-7,其余种质介于两者之间。因此,本研究筛选出苗期抗旱性强的薏苡种质分别为yy18-1、yy14-3和yy13-1,可为薏苡抗旱育种、抗旱机理及干旱调控缓解机制的研究提供基础材料。

3.3 薏苡种质资源苗期抗旱指标的筛选

作物的抗旱性是复杂的数量性状,是众多因素、多种机制共同作用的结果,最终通过各种指标在不同生育时期的一系列变化表现出来[22]。因此,指标的合理选择是作物抗旱性鉴定的关键。目前,国内外****在作物苗期抗旱指标筛选方面开展了大量研究工作,并针对不同的作物筛选出了不同的抗旱指标[12-18]。本研究中,由于各指标受干旱胁迫影响的程度各异,且各指标间存在一定程度的相关性。因此,直接利用这些指标很难客观、准确地评价各种质的抗旱性,从而影响抗旱鉴定结果。各指标与D值的密切程度与各指标对干旱胁迫反应的敏感性及各指标与WDC值的密切程度吻合。通过逐步回归分析,得到与D值密切相关的指标有幼苗干旱存活率、株高、叶宽、根长和根鲜重。因此,幼苗干旱存活率、株高、叶宽、根长和根鲜重可作为薏苡种质资源苗期鉴定、直观的抗旱性评价指标。

4 结论

干旱胁迫对薏苡种质资源苗期各指标均有极显著影响。苗期抗旱性强的薏苡种质分别为yy18-1、yy14-3和yy13-1,可为薏苡抗旱育种、抗旱机理及干旱调控缓解机制的研究提供基础材料。幼苗存活率、株高、叶宽、根长和根鲜重可作为评价薏苡种质资源苗期抗旱性的指标性状。
Attached table 1
附表1
附表150份薏苡种质信息
Attached table 1The information of 50 Job’s tears germplasms
编号
Number
名称
Name
来源
Origin
编号
Number
名称
Name
来源
Origin
YG01yy13-1野生种
Wild species
YG26yy03-4晴隆地方品种
Local cultivar in Qianglong
YG02yy13-2野生种
Wild specie
YG27yy18-1兴仁地方品种
Local cultivar in Xingren
YG03yy13-3野生种
Wild specie
YG28yy12-2正安地方品种
Local cultivar in Zhengan
YG04粱丰薏16-2
Liiangfengyi 16-2
黔薏苡2号选系
Derived from Qianyiyi 2
YG29yy14-5广西地方品种
Local cultivar in Guangxi
YG05黔薏苡1号
Qianyiyi 1
国审品种
National authorized cultivar
YG30yy08-4安龙地方品种
Local cultivar in Anlong
YG06黔薏苡16-1
Qianyiyi 16-1
黔薏苡1号选系
Derived from Qianyiyi 1
YG31yy14-2广西地方品种
Local cultivar in Guangxi
YG07粱丰薏16-1
Liiangfengyi 16-1
黔薏苡1号选系
Derived from Qianyiyi 1
YG32yy16-3兴仁地方品种
Local cultivar in Xingren
YG08黔薏苡16-2
Qianyiyi 16-2
黔薏苡2号选系
Derived from Qianyiyi 2
YG33yy07-6盘县地方品种
Local cultivar in Panxian
YG09粱丰薏14-1
Liangfengyi 14-1
黔薏苡1号选系
Derived from Qianyiyi 1
YG34yy14-6广西地方品种
Local cultivar in Guangxi
YG10粱丰薏14-2
Liangfengyi 14-2
黔薏苡2号选系
Derived from Qianyiyi 2
YG35yy03-2晴隆地方品种
Local cultivar in Qianglong
YG11yy07-8盘县地方品种
Local cultivar in Panxian
YG36yy03-8晴隆地方品种
Local cultivar in Qianglong
YG12yy12-1正安地方品种
Local cultivar in Zhengan
YG37yy14-3四川地方品种
Local cultivar in Sichuan
YG13yy04-2云南地方品种
Local cultivar in Yunnan
YG38yy14-10四川地方品种
Local cultivar in Sichuan
YG14yy07-2盘县地方品种
Local cultivar in Panxian
YG39yy03-7晴隆地方品种
Local cultivar in Qianglong
YG15yy04-7云南地方品种
Local cultivar in Yunnan
YG40yy14-7四川地方品种
Local cultivar in Sichuan
YG16yy07-3盘县地方品种
Local cultivar in Panxian
YG41yy07-7盘县地方品种
Local cultivar in Panxian
YG17yy03-6晴隆地方品种
Local cultivar in Qianglong
YG42yy06-1兴仁地方品种
Local cultivar in Xingren
YG18yy07-5盘县地方品种
Local cultivar in Panxian
YG43yy07-10盘县地方品种
Local cultivar in Panxian
YG19yy11-2广西地方品种
Local cultivar in Guangxi
YG44yy12-3正安地方品种
Local cultivar in Zhengan
YG20yy07-1盘县地方品种
Local cultivar in Panxian
YG45yy04-6云南地方品种
Local cultivar in Yunnan
YG21黔薏苡2号
Qianyiyi 2
国审品种
National authorized cultivar
YG46yy19-8安龙地方品种
Local cultivar in Anlong
YG22yy11-8广西地方品种
Local cultivar in Guangxi
YG47yy18-4兴仁地方品种
Local cultivar in Xingren
YG23yy08-9安龙地方品种
Local cultivar in Anlong
YG48yy08-5安龙地方品种
Local cultivar in Anlong
YG24yy18-2兴仁地方品种
Local cultivar in Xingren
YG49yy07-9盘县地方品种
Local cultivar in Panxian
YG25yy07-4盘县地方品种
Local cultivar in Panxian
YG50yy12-7正安地方品种
Local cultivar in Zhengan


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The authors have declared that no competing interests exist.

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