Accumulation and Molecular Detection of Pb and Cd in Sweet Corn Inbred Lines
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摘要:
目的 筛选获得在重金属Pb、Cd单一污染及复合污染下Pb、Cd低积累的优良甜玉米自交系，以及可用于甜玉米重金属Pb、Cd积累早代鉴定的分子技术。 方法 以10个甜玉米自交系为供试材料，通过盆栽试验研究重金属Pb、Cd单一污染及Pb、Cd复合污染下，玉米根系、茎叶和籽粒中Pb、Cd含量的积累差异；同时对供试材料ZmHMA2 InDel位点进行Pb、Cd积累差异分子检测，综合分子标记检测与重金属Pb、Cd在不同材料间的积累结果，筛选甜玉米重金属低积累自交系。 结果 无论是单一污染还是复合污染，重金属Pb、Cd的积累规律均表现为根系＞茎叶＞籽粒；在复合污染下，玉米不同组织对重金属Pb、Cd的积累无明显竞争与协同效应，表明玉米Pb、Cd的积累机制存在一定差异。通过鉴定，获得籽粒Pb低积累玉米自交系2份，籽粒Cd低积累玉米自交系3份，仅有一份材料闽甜系X901表现出籽粒Pb、Cd均低积累。利用InDel 位点（InDel2307）对供试材料进行分子检测，结果发现供试材料中有4份存在该位点，含有该位点的材料中根系、茎叶和籽粒Cd含量平均值较其他材料平均值分别低1.801、0.64 、0.131 mg·kg−1。 结论 InDel2307位点能将不同玉米自交系按Cd累积量进行区分，对甜玉米Cd含量的区分具有特异性标记。综合分子标记检测与Pb、Cd含量积累结果，筛选出自交系闽甜系X901 为Pb、Cd低积累材料。 Abstract:Objective Sweet corn inbred lines low in accumulating Pb and Cd were identified, and a molecular detection method for the pollutants examined. Method Ten sweet corn inbred lines were tested on Pb and Cd accumulation. In a pot experiment, the heavy metals in roots, stems/leaves, and kernels of the plants grown on soil with spiked Pb and/or Cd were measured over time. Applicability of a molecular detection method on Pb and Cd at the InDel locus of ZmHMA2 in corn plant was scrutinized. Result The accumulation of Pb and Cd in organs of a corn plant was roots＞stems/leaves＞kernels. It did not differ significantly whether the metal elements were presented individually or simultaneously in the soil. Two inbred lines were identified to retain in the kernels less on Pb, 3 on Cd, and only Mintian X901 on both Pb and Cd. The targeted molecular detection found InDel 2307 locus in 4 of the specimens, the average content of Cd which contained were 1.801 mg·kg−1 lower in the roots, 0.64 mg·kg−1 lower in the stems/leaves, and 0.131 mg·kg−1 lower in the kernels compared to the average of the other varieties. Conclusion The InDel 2307 locus as a marker displayed a specificity in differentiating Cd content in corn plants grown on soils contaminated with different levels of the heavy metals. The molecular detection result and the Pb and Cd contents on the 10 sweet corn inbred lines indicated that Mintian X901 was the cultivar least prone to accumulate the pollutants. -
Key words:
- Heavy metal /
- Pb pollution /
- Cd pollution /
- sweet corn /
- accumulation /
- marker detection
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图 1 10个玉米自交系Indel2307位点的序列比对
Figure 1. Sequence alignment of Indel 2307 locus in 10 sweet corn inbred lines
表 1 供试甜玉米材料编号及名称
Table 1. Codes and names of sweet corn inbred lines
编号
No.自交系
Inbred line编号
No.自交系
Inbred lineS1 闽甜系688 S6 闽甜系AS74 S2 闽甜系G23 S7 闽甜系AS76 S3 闽甜系H6 S8 闽甜系T146 S4 闽甜系197 S9 闽甜系X901 S5 闽甜系AS67 S10 闽甜系JR8609 
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表 2 ZmHMA2基因所用引物
Table 2. Primers used in amplifying ZmHMA2
引物名称
Primer name正向引物 (5′-3′)
Primer sequence(5′-3′)反向引物(5′-3′)
Primer sequence(5′-3′)ZmHMA2-1 TTATCGCTCCGGATATGCCC CCATCCATCCCTTCAGCCTTT ZmHMA2-2 GGATATGCCCTCTCCAGGGT CCATCCATCCCTTCAGCCTT ZmHMA2-3 TTTATCGCTCCGGATATGCCC CATCCATCCCTTCAGCCTTT
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表 3 单一Pb污染环境下玉米不同基因型及不同部位重金属Pb含量
Table 3. Pb contents in tissues of sweet corn inbred lines grown under Pb-contaminated soil
自交系
Inbred linePb含量 Pb content/（mg·kg−1） 分子检测
Marker detection根 Roots 茎叶 Shoots 籽粒 Grains 闽甜系688(S1) 240.171±1.023 bcA 89.531±1.101 cB 0.312±0.019 bC − 闽甜系G23(S2) 214.412±1.201 dA 87.408±1.062 cB 0.244±0.018 cC − 闽甜系H6(S3) 176.303±2.390 efA 86.721±1.112 cB 0.135±0.018 eC − 闽甜系197(S4) 233.591±2.331 cA 87.295±0.945 cB 0.223±0.009 cC + 闽甜系AS67(S5) 259.374±3.031 aA 125.216±2.213 aB 0.381±0.010 aC + 闽甜系AS74(S6) 249.845±1.944 bA 120.911±1.409 abB 0.321±0.011 a C + 闽甜系AS76(S7) 248.210±2.802 bA 90.923±1.204 cB 0.227±0.017 cC − 闽甜系T146(S8) 260.183±1.255 aA 114.475±1.301 bB 0.397±0.018 aC − 闽甜系X901(S9) 169.003±3.501 fA 89.782±0.928 cB 0.172±0.010 dC + 闽甜系JR8609(S10) 185.784±2.902 eA 91.761±2.691 cB 0.344±0.010 bC − 同行数据后不同大写字母表示不同组织部位间差异极显著（P＜0.01），同列数据后不同小写字母表示不同自交系间差异显著（P＜0.05）。下表同。
Data with different capital letters on same row indicate extremely significant differences among different tissues at P＜0.01; those with different lowercase letters on the same column indicate significant differences among different inbred line at P＜0.05. Same for below.
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表 4 单一Cd污染环境下玉米不同基因型及不同部位重金属Cd含量
Table 4. Cd contents in tissues of sweet corn inbred lines grown under Cd-contaminated soil
编号
No.Cd含量 Cd content/（mg·kg−1） 根系 Roots 茎叶 Shoots 籽粒 Grains 闽甜系688(S1) 3.528±0.102 aA 1.723±0.116 abB 0.213±0.121 bC 闽甜系G23(S2) 3.823±0.123 aA 1.285±0.126 bcB 0.181±0.120 cC 闽甜系H6(S3) 3.124±0.022 bA 1.992±0.092 aB 0.241±0.018 aC 闽甜系197(S4) 1.055±0.134 eA 1.137±0.201 cB 0.106±0.015 dC 闽甜系AS67(S5) 1.231±0.094 eA 0.988±0.132 dB 0.086±0.021 eC 闽甜系AS74(S6) 1.166±0.236 eA 0.932±0.144 dB 0.091±0.012 deC 闽甜系AS76(S7) 2.931±0.136 cA 2.022±0.118 aB 0.234±0.011 aC 闽甜系T146(S8) 2.215±0.254 cdA 1.535±0.087 bB 0.209±0.011 bC 闽甜系X901(S9) 1.043±0.108 eA 0.772±0.102 eB 0.063±0.001 fC 闽甜系JR8609(S10) 1.929±0.203 dA 1.025±0.121 dB 0.225±0.012 abC
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表 5 Pb-Cd复合污染下玉米不同基因型及不同部位重金属Pb、Cd含量
Table 5. Pb and Cd contents in tissues of sweet corn inbred lines grown under Pb-and-Cd-contaminated soil
编号
No.根系重金属含量
Heavy metal content in roots/(mg·kg−1)茎叶重金属含量
Heavy metal content in shoots/(mg·kg−1)籽粒重金属含量
Heavy metal content in grains/(mg·kg−1)Pb Cd Pb Cd Pb Cd 闽甜系688(S1) 225.182±2.211 c 2.981±0.130 a 95.427±2.122 b 1.231±0.122 c 0.432±0.022 ab 0.360±0.020 a 闽甜系G23(S2) 218.523±4.652 d 2.594±0.221 b 94.143±1.842 b 0.924±0.114 c 0.392±0.021 b 0.141±0.021 c 闽甜系H6(S3) 167.623±3.692 f 3.049±0.182 a 86.613±1.553 c 1.771±0.191 a 0.083±0.038 e 0.350±0.025 a 闽甜系197(S4) 219.825±2.622 d 1.911±0.372 c 95.310±1.190 b 0.908±0.127 d 0.309±0.011 c 0.091±0.024 d 闽甜系AS67(S5) 234.535±4.741 b 1.263±0.218 d 115.542±1.424 a 0.815±0.102 d 0.429±0.017 ab 0.087±0.019 d 闽甜系AS74(S6) 228.191±3.505 c 2.406±0.132 b 119.261±1.211 a 1.197±0.119 c 0.471±0.022 a 0.102±0.019 c 闽甜系AS76(S7) 228.435±2.142 c 3.044±0.159 a 79.243±2.107 d 1.551±0.149 b 0.383±0.033 b 0.178±0.022 b 闽甜系T146(S8) 246.653±2.712 a 1.951±0.154 c 111.020±1.508 a 1.168±0.214 c 0.463±0.018 a 0.155±0.024 bc 闽甜系X901(S9) 159.371±3.443 f 1.221±0.213 d 93.035±1.416 b 0.536±0.275 e 0.094±0.018 e 0.074±0.020 d 闽甜系JR8609(S10) 196.029±1.961 e 1.408±0.126 d 85.861±1.312 c 0.860±0.112 d 0.245±0.032 d 0.151±0.021 c
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