两种叶菜镉、铅低积累品种筛选及其快速鉴别方法研究
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摘要
以受重金属污染较为严重的芹菜(Apium graveolens L.))和油麦菜(Lactuca dolichophylla Kitam.)为研究对象,通过盆栽试验,研究了27个芹菜品种和28个油麦菜品种吸收积累Cd、Pb的品种间差异,对Cd、Pb的转运能力,茎叶积累Cd、Pb的稳定性及Cd+Pb-PSC的筛选(Pollution-safe cultivar,指在一定污染水平的土壤中作物可食部分吸收积累污染物含量符合食品安全标准、并且经过验证确认其污染物低量积累特性是稳定的一类品种,简称PSC)。以本研究和前期试验筛选到的不同蔬菜Cd积累典型品种为研究对象,研究了其市售种子中Cd含量和植物发育初期幼苗中NaCl提取态、稀HCl提取态Cd含量在同种蔬菜不同Cd积累能力品种间的差异。利用基因克隆和基因测序技术,研究了10个油麦菜Cd积累典型品种LdPCS1基因单核苷酸多态性(single nucleotide polymorphism, SNP)。在此基础上,探讨了叶用蔬菜Cd低积累基因型的快速筛选方法。主要研究结果如下:
一、通过盆栽试验研究了3个土壤Cd含量(L :0.142 mg kg(-1),M:0.431 mg kg(-1);H:0.631 mg kg(-1))下27个芹菜品种对Cd积累的品种间差异,对供试芹菜品种茎叶生物量和Cd含量的二元方差分析表明,由品种、土壤Cd处理以及品种×土壤Cd处理间交互作用引起的茎叶生物量和Cd含量变异均达到极显著水平(p<0.01),茎叶Cd含量(基于鲜重)在品种间的变异范围分别为0.046 - 0.218 mg kg(-1)、0.270 - 0.633 mg kg(-1)和0.208 - 0.686 mg kg(-1),变异系数分别为26.9%、20.9%和24.5%。当土壤Cd含量为0.142mg kg(-1)时,有1个品种超出国际食品法典委员会(CAC)标准中叶菜Cd含量最高限值(0.2mg kg(-1) FW),超标率3.7%,当土壤Cd含量为0.431mg kg(-1)和0.631mg kg(-1)时,供试芹菜品种茎叶Cd含量全部超标。因此,当土壤Cd含量> 0.431mg kg(-1)时不存在Cd-PSC,说明芹菜是非常容易受Cd污染的蔬菜种类。而有8个品种(No.7、9、20、22、24、25、26、28)具有相对较低的茎叶Cd含量及较大的生物量,可作为典型Cd低积累品种。No.2可作为Cd高积累品种。
二、通过盆栽试验研究了芹菜8个Cd低积累品种和1个Cd高积累品种(No.2)对Cd、Pb胁迫响应的品种间差异,对供试芹菜品种茎叶Cd、Pb含量的二元方差分析表明,由品种、Cd或Pb处理以及品种×Cd或品种×Pb处理的交互作用引起的变异均达到显著水平(p<0.05),在土壤Cd含量为0.35 mg kg(-1),Pb含量为49.89 mg kg(-1)时,5个品种(No.7、9、22、24、25)可作为Cd-PSCs、Pb-PSCs及Cd+Pb-PSCs。其中No.25在高Cd胁迫下(土壤Cd含量0.599~0.667 mg kg(-1)),其茎叶Cd含量仅是No.2(Cd高积累品种)茎叶Cd含量的41.93%~59.70%,其茎叶Pb含量在土壤Pb含量高达231 mg kg(-1)时能够接近CAC标准的Pb含量最大限值,可作为适合于农业推广的芹菜Cd+Pb-PSC品种,可有效降低Cd、Pb对芹菜的污染。
三、通过盆栽试验研究了3个土壤Cd含量下(L :0.129 mg kg(-1),M:0.352 mg kg(-1);H:1.253 mg kg(-1))下28个油麦菜品种对Cd积累的品种间差异,对供试油麦菜品种茎叶生物量和Cd含量的二元方差分析表明,由品种、土壤Cd处理以及品种×土壤Cd处理间交互作用引起的茎叶生物量和Cd含量变异均达到极显著水平(p<0.01),茎叶Cd含量(基于鲜重)在品种间的变异范围分别为0.027 - 0.140 mg kg(-1)、0.084 - 0.400 mg kg(-1)和0.358 - 1.715 mg kg(-1),变异系数分别为38.5%、36.1%和46.5%。当土壤Cd含量为0.129mg kg(-1)时,供试油麦菜品种茎叶Cd含量全部符合国际食品法典委员会(CAC)标准中叶菜Cd含量最高限值(0.2mg kg(-1) FW)。当土壤Cd含量为0.352mg kg(-1)时,有13个品种的茎叶Cd含量低于0.2 mg kg(-1) FW,超标率为53.6%。当土壤Cd含量为1.253 mg kg(-1)时,供试油麦菜品种茎叶Cd含量全部超标。可见,当土壤Cd含量> 0.352mg kg(-1)时不存在Cd-PSC,6个品种(No.3、6、7、8、10、11)具有相对较低的茎叶Cd含量及较大的生物量,可作为Cd-PSCs,而No.19、23、24具有较高的茎叶Cd含量和较大的生物量,可作为Cd高积累品种用于后继研究。
四、通过盆栽试验研究了油麦菜6个Cd低积累品种和1个Cd高积累品种(No.19)对Cd、Pb胁迫响应的品种间差异,对供试油麦菜品种茎叶Cd、Pb含量的二元方差分析表明,由品种、Cd或Pb处理以及品种×Cd或品种×Pb处理的交互作用引起的变异均达到极显著水平(p<0.01),在Cd含量小于0.352 mg kg(-1),Pb含量小于247 mg kg(-1)的Cd、Pb污染土壤中,品种No.7、11、8、10可以作为Cd+Pb-PSCs。
五、分析了前期筛选得到的芹菜、油麦菜、蕹菜、苋菜、菜心Cd积累典型品种市售种子中Cd含量,结果表明,芹菜市售种子中Cd含量在Cd高积累和低积累品种间差异十分明显,且与盆栽试验中地上部Cd含量的品种间差异间有显著相关性,因此,市售种子中Cd含量可以用于芹菜Cd低积累品种的快速筛选。
六、水培条件下对前期筛选得到的油麦菜、蕹菜、小白菜、菜心Cd积累典型品种发育初期幼苗中NaCl提取态和稀HCl提取态Cd含量研究发现,利用NaCl提取态Cd含量与稀HCl提取态Cd含量的比值作为衡量植物积累Cd能力的指标,对小白菜和蕹菜而言,能够十分有效地区分Cd低积累和高积累品种,对其他种类蔬菜有待进一步研究。七、对10个油麦菜Cd积累典型品种Cd积累相关基因LdPSC1的单核苷酸
多态性(SNP)研究发现,Cd高积累品种与低积累品种间存在特异性的突变位点。中性检验结果显示Cd高积累品种与低积累品种间有分化的趋势,该基因在Cd胁迫下经历强烈的净化选择(negative selection)作用。研究结果为今后进一步探讨Cd低积累品种的分子标记方法,发展分子辅助育种技术提供理论依据。
This study take celery (Apium graveolens L.) and lettuce (Lactuca dolichophylla Kitam.) that are vulnerable to contamination by soil Cd and Pb as the object. Two pot experiments were conducted to investigate the variations, transfer potential, and stability of Cd and Pb accumulations in 27 celery genotypes and 28 lettuce genotypes respectively, and to screen for Cd+Pb-PSC (pollution-safe cultivar , which edible parts accumulate certain pollutant at low enough level for safe consumption while grown in contaminated soil). Using vegetable typical Cd accumulative genotypes that got from this study and preliminary experiments, variations of their Cd concentration of commercial seed, NaCl extractable or HCl extractable Cd concentration in their plant seedlings in early development were investigated. Using gene cloning and gene sequencing technology,the research of single nucleotide polymorphism in LdPCS1 gene in 10 typical Cd accumulative genotypes of lettuce were carried out. On this basis, we set up the rapid screening methods for low Cd accumulative genotype of leaf vegetables. Main results gained are summarized as follows: 1. In order to investigate the genotypic variations in accumulation of Cd in celery,
a pot experiment was conducted on 27 celery cultivars under three soil Cd levels (L: 0.142 mg kg~(-1),M: 0.431 mg kg~(-1);H: 0.631 mg kg~(-1)). Results of two-way ANOVA for the variations of Cd concentrations and biomass in shoots showed that variations from genotypes, soil Cd and interaction of genotype×soil Cd were all significant (p < 0.01), the Cd concentration (Fresh basis)in the shoot ranged from 0.046 to 0.218 mg kg~(-1), 0.270 to 0.633 mg kg~(-1) and 0.208 to 0.686 mg kg~(-1), their coefficient of variation (CV) were 26.9%, 20.9% and 24.5% under soil Cd treatments L, M and H, respectively. In treatment L (soil Cd 0.142 mg kg~(-1)), except for one, shoot Cd concentrations in almost all tested cultivars in treatment L were lower than the ML of Cd allowed by CAC standards (0.2 mg kg~(-1) FW), 3.7% tested cultivars being higher than the limitation. However, none of the tested cultivars measured up to the standard in treatment M and treatment H (soil Cd 0.431 mg kg~(-1) and 0.631 mg kg~(-1), respectively). that was to say, there was no Cd-PSC among the tested cultivars when soil Cd concentration > 0.431 mg kg~(-1). This suggests that celery is a vegetable being easily polluted by Cd in soil. These eight genotypes (No.7、9、20、22、24、25、26、28) with lower shoot Cd concentrations and higher shoot biomass, could be treated as typical low-Cd accumulating cultivars. The genotype (No.2) could be treated as a typical high-Cd accumulating cultivar.
2. The other pot experiment was carried out to study the differences of accumulation of Cd and Pb by these typical Cd accumulative cultivars. variations of Cd and Pb concentrations in shoot from genotypes, soil Cd or Pb level and genotype×soil Cd or Pb level were all significant at p < 0.05 level, In treatment LCdLPb (soil Cd 0.142 mg kg~(-1), Pb 49.89 mg kg~(-1)), five cultivars (No.7、9、22、24、25) can be used as Cd-PSCs, Pb-PSCs and Cd+Pb-PSCs. Among them, shoot Cd concentration of No.25 is only 41.93%~59.70% of No.2 under high Cd exposure (soil Cd 0.599~0.667 mg kg~(-1)), and shoot Pb concentration of No.25 near the ML for Pb allowed by the CAC standards. The genotype No.25 is identified as adoptable Cd+Pb-PSC which is worth to recommend to growers, and adopting PSC strategy could help reduce the risk of Cd, Pb pollution in celery.
3. A pot experiment was carried out to investigate the genotypic variations in accumulation of Cd in lettuce under three soil Cd treatment (L: 0.129 mg kg~(-1),M: 0.352 mg kg~(-1);H: 1.253 mg kg~(-1)). Results of two-way ANOVA for the variations of Cd concentrations and biomass in shoots showed that variations from genotypes, soil Cd and interaction of genotype×soil Cd were all significant (p < 0.01), the Cd concentration (Fresh basis)in the shoot ranged from 0.027 to 0.140 mg kg~(-1), 0.084 to 0.400 mg kg~(-1) and 0.358 to 1.715 mg kg~(-1), their coefficient of variation (CV) were 38.5%, 36.1% and 46.5% under soil Cd treatments L, M and H, respectively. In treatment L (soil Cd 0.129 mg kg~(-1)), shoot Cd concentrations in all tested cultivars were lower than the ML of Cd allowed by CAC standards (0.2 mg kg~(-1) FW). Thirteen of the tested cultivars measured up to the standard in treatment M (soil Cd 0.352 mg kg~(-1)), 53.6% tested cultivars being higher than the limitation. However, none of the tested cultivars measured up to the standard in treatment treatment H (soil Cd 1.253 mg kg~(-1)). Thus, no Cd-PSC could be found when soil Cd concentration higher than 0.352 mg kg~(-1). only 6 cultivars(No.3、6、7、8、10、11)with lower shoot Cd concentrations and higher shoot biomasses, could be treated as Cd-PSCs, and 3 cultivars(No.19、23、24)with higher shoot Cd concentrations and higher shoot biomasses, could be treated as non-Cd-PSC in the follow-up study.
4. The other pot experiment was carried out to study the variations of accumulation of Cd and Pb by six Cd-PSCs and one non-Cd-PSC. Variations of Cd and Pb concentrations in shoot from genotypes, soil Cd or Pb level and genotype×soil Cd or Pb level were all significant at p < 0.01 level, under lower 0.352 mg kg~(-1)soil Cd level and lower 247 mg kg~(-1)soil Pb level, 4 cultivars(No.7、8、10、11)could be treated as Cd+Pb-PSCs.
5. Detected the seeds Cd concentration in typical Cd accumulation cultivars of celery, lettuce, water spinach, amaranth and Chinese flowering cabbage that screened in early experiments, the results show that, the significant differences were observed between low-Cd Cultivars and high-Cd cultivars of celery, and the seeds Cd concentration of the tested cultivars of celery was significantly correlated with those shoot Cd concentration in pot experiment. So we can use the seed Cd concentration to rapid identify the low-Cd cultivars of celery.
6. Sequential extraction method was used to analyze and distinguish NaCl extractable forms and HCl extractable forms of Cd in seedlings of typical Cd accumulation cultivars of lettuce, water spinach, Chinese cabbage and Chinese flowering cabbage that screened in early experiments, exposed to Cd. The studies showed that, it can be very effective distinguish low-Cd and high-Cd accumulative cultivars for Chinese cabbage and water spinach by using the ratio value of NaCl extractable Cd : HCl extractable Cd. But for the other vegetable species, it should be studied further.
7. In an effort to understand the single nucleotide polymorphisms (SNP) in LdPSC1 gene in 10 typical Cd accumulative cultivars of lettuce, we discovered the specific mutations existed between low-Cd cultivars and high-Cd cultivars in LdPSC1 gene. The test of departure of neutral mutation of nucleotide variation in LdPSC1 gene showed there was a tendency to divide in low-Cd cultivars and high-Cd cultivars, the LdPSC1 gene experiencing strong negative selection under Cd stress. The results could provide a theoretical basis for using the molecular markers in breeding of low-Cd cultivars.
一、通过盆栽试验研究了3个土壤Cd含量(L :0.142 mg kg(-1),M:0.431 mg kg(-1);H:0.631 mg kg(-1))下27个芹菜品种对Cd积累的品种间差异,对供试芹菜品种茎叶生物量和Cd含量的二元方差分析表明,由品种、土壤Cd处理以及品种×土壤Cd处理间交互作用引起的茎叶生物量和Cd含量变异均达到极显著水平(p<0.01),茎叶Cd含量(基于鲜重)在品种间的变异范围分别为0.046 - 0.218 mg kg(-1)、0.270 - 0.633 mg kg(-1)和0.208 - 0.686 mg kg(-1),变异系数分别为26.9%、20.9%和24.5%。当土壤Cd含量为0.142mg kg(-1)时,有1个品种超出国际食品法典委员会(CAC)标准中叶菜Cd含量最高限值(0.2mg kg(-1) FW),超标率3.7%,当土壤Cd含量为0.431mg kg(-1)和0.631mg kg(-1)时,供试芹菜品种茎叶Cd含量全部超标。因此,当土壤Cd含量> 0.431mg kg(-1)时不存在Cd-PSC,说明芹菜是非常容易受Cd污染的蔬菜种类。而有8个品种(No.7、9、20、22、24、25、26、28)具有相对较低的茎叶Cd含量及较大的生物量,可作为典型Cd低积累品种。No.2可作为Cd高积累品种。
二、通过盆栽试验研究了芹菜8个Cd低积累品种和1个Cd高积累品种(No.2)对Cd、Pb胁迫响应的品种间差异,对供试芹菜品种茎叶Cd、Pb含量的二元方差分析表明,由品种、Cd或Pb处理以及品种×Cd或品种×Pb处理的交互作用引起的变异均达到显著水平(p<0.05),在土壤Cd含量为0.35 mg kg(-1),Pb含量为49.89 mg kg(-1)时,5个品种(No.7、9、22、24、25)可作为Cd-PSCs、Pb-PSCs及Cd+Pb-PSCs。其中No.25在高Cd胁迫下(土壤Cd含量0.599~0.667 mg kg(-1)),其茎叶Cd含量仅是No.2(Cd高积累品种)茎叶Cd含量的41.93%~59.70%,其茎叶Pb含量在土壤Pb含量高达231 mg kg(-1)时能够接近CAC标准的Pb含量最大限值,可作为适合于农业推广的芹菜Cd+Pb-PSC品种,可有效降低Cd、Pb对芹菜的污染。
三、通过盆栽试验研究了3个土壤Cd含量下(L :0.129 mg kg(-1),M:0.352 mg kg(-1);H:1.253 mg kg(-1))下28个油麦菜品种对Cd积累的品种间差异,对供试油麦菜品种茎叶生物量和Cd含量的二元方差分析表明,由品种、土壤Cd处理以及品种×土壤Cd处理间交互作用引起的茎叶生物量和Cd含量变异均达到极显著水平(p<0.01),茎叶Cd含量(基于鲜重)在品种间的变异范围分别为0.027 - 0.140 mg kg(-1)、0.084 - 0.400 mg kg(-1)和0.358 - 1.715 mg kg(-1),变异系数分别为38.5%、36.1%和46.5%。当土壤Cd含量为0.129mg kg(-1)时,供试油麦菜品种茎叶Cd含量全部符合国际食品法典委员会(CAC)标准中叶菜Cd含量最高限值(0.2mg kg(-1) FW)。当土壤Cd含量为0.352mg kg(-1)时,有13个品种的茎叶Cd含量低于0.2 mg kg(-1) FW,超标率为53.6%。当土壤Cd含量为1.253 mg kg(-1)时,供试油麦菜品种茎叶Cd含量全部超标。可见,当土壤Cd含量> 0.352mg kg(-1)时不存在Cd-PSC,6个品种(No.3、6、7、8、10、11)具有相对较低的茎叶Cd含量及较大的生物量,可作为Cd-PSCs,而No.19、23、24具有较高的茎叶Cd含量和较大的生物量,可作为Cd高积累品种用于后继研究。
四、通过盆栽试验研究了油麦菜6个Cd低积累品种和1个Cd高积累品种(No.19)对Cd、Pb胁迫响应的品种间差异,对供试油麦菜品种茎叶Cd、Pb含量的二元方差分析表明,由品种、Cd或Pb处理以及品种×Cd或品种×Pb处理的交互作用引起的变异均达到极显著水平(p<0.01),在Cd含量小于0.352 mg kg(-1),Pb含量小于247 mg kg(-1)的Cd、Pb污染土壤中,品种No.7、11、8、10可以作为Cd+Pb-PSCs。
五、分析了前期筛选得到的芹菜、油麦菜、蕹菜、苋菜、菜心Cd积累典型品种市售种子中Cd含量,结果表明,芹菜市售种子中Cd含量在Cd高积累和低积累品种间差异十分明显,且与盆栽试验中地上部Cd含量的品种间差异间有显著相关性,因此,市售种子中Cd含量可以用于芹菜Cd低积累品种的快速筛选。
六、水培条件下对前期筛选得到的油麦菜、蕹菜、小白菜、菜心Cd积累典型品种发育初期幼苗中NaCl提取态和稀HCl提取态Cd含量研究发现,利用NaCl提取态Cd含量与稀HCl提取态Cd含量的比值作为衡量植物积累Cd能力的指标,对小白菜和蕹菜而言,能够十分有效地区分Cd低积累和高积累品种,对其他种类蔬菜有待进一步研究。七、对10个油麦菜Cd积累典型品种Cd积累相关基因LdPSC1的单核苷酸
多态性(SNP)研究发现,Cd高积累品种与低积累品种间存在特异性的突变位点。中性检验结果显示Cd高积累品种与低积累品种间有分化的趋势,该基因在Cd胁迫下经历强烈的净化选择(negative selection)作用。研究结果为今后进一步探讨Cd低积累品种的分子标记方法,发展分子辅助育种技术提供理论依据。
This study take celery (Apium graveolens L.) and lettuce (Lactuca dolichophylla Kitam.) that are vulnerable to contamination by soil Cd and Pb as the object. Two pot experiments were conducted to investigate the variations, transfer potential, and stability of Cd and Pb accumulations in 27 celery genotypes and 28 lettuce genotypes respectively, and to screen for Cd+Pb-PSC (pollution-safe cultivar , which edible parts accumulate certain pollutant at low enough level for safe consumption while grown in contaminated soil). Using vegetable typical Cd accumulative genotypes that got from this study and preliminary experiments, variations of their Cd concentration of commercial seed, NaCl extractable or HCl extractable Cd concentration in their plant seedlings in early development were investigated. Using gene cloning and gene sequencing technology,the research of single nucleotide polymorphism in LdPCS1 gene in 10 typical Cd accumulative genotypes of lettuce were carried out. On this basis, we set up the rapid screening methods for low Cd accumulative genotype of leaf vegetables. Main results gained are summarized as follows: 1. In order to investigate the genotypic variations in accumulation of Cd in celery,
a pot experiment was conducted on 27 celery cultivars under three soil Cd levels (L: 0.142 mg kg~(-1),M: 0.431 mg kg~(-1);H: 0.631 mg kg~(-1)). Results of two-way ANOVA for the variations of Cd concentrations and biomass in shoots showed that variations from genotypes, soil Cd and interaction of genotype×soil Cd were all significant (p < 0.01), the Cd concentration (Fresh basis)in the shoot ranged from 0.046 to 0.218 mg kg~(-1), 0.270 to 0.633 mg kg~(-1) and 0.208 to 0.686 mg kg~(-1), their coefficient of variation (CV) were 26.9%, 20.9% and 24.5% under soil Cd treatments L, M and H, respectively. In treatment L (soil Cd 0.142 mg kg~(-1)), except for one, shoot Cd concentrations in almost all tested cultivars in treatment L were lower than the ML of Cd allowed by CAC standards (0.2 mg kg~(-1) FW), 3.7% tested cultivars being higher than the limitation. However, none of the tested cultivars measured up to the standard in treatment M and treatment H (soil Cd 0.431 mg kg~(-1) and 0.631 mg kg~(-1), respectively). that was to say, there was no Cd-PSC among the tested cultivars when soil Cd concentration > 0.431 mg kg~(-1). This suggests that celery is a vegetable being easily polluted by Cd in soil. These eight genotypes (No.7、9、20、22、24、25、26、28) with lower shoot Cd concentrations and higher shoot biomass, could be treated as typical low-Cd accumulating cultivars. The genotype (No.2) could be treated as a typical high-Cd accumulating cultivar.
2. The other pot experiment was carried out to study the differences of accumulation of Cd and Pb by these typical Cd accumulative cultivars. variations of Cd and Pb concentrations in shoot from genotypes, soil Cd or Pb level and genotype×soil Cd or Pb level were all significant at p < 0.05 level, In treatment LCdLPb (soil Cd 0.142 mg kg~(-1), Pb 49.89 mg kg~(-1)), five cultivars (No.7、9、22、24、25) can be used as Cd-PSCs, Pb-PSCs and Cd+Pb-PSCs. Among them, shoot Cd concentration of No.25 is only 41.93%~59.70% of No.2 under high Cd exposure (soil Cd 0.599~0.667 mg kg~(-1)), and shoot Pb concentration of No.25 near the ML for Pb allowed by the CAC standards. The genotype No.25 is identified as adoptable Cd+Pb-PSC which is worth to recommend to growers, and adopting PSC strategy could help reduce the risk of Cd, Pb pollution in celery.
3. A pot experiment was carried out to investigate the genotypic variations in accumulation of Cd in lettuce under three soil Cd treatment (L: 0.129 mg kg~(-1),M: 0.352 mg kg~(-1);H: 1.253 mg kg~(-1)). Results of two-way ANOVA for the variations of Cd concentrations and biomass in shoots showed that variations from genotypes, soil Cd and interaction of genotype×soil Cd were all significant (p < 0.01), the Cd concentration (Fresh basis)in the shoot ranged from 0.027 to 0.140 mg kg~(-1), 0.084 to 0.400 mg kg~(-1) and 0.358 to 1.715 mg kg~(-1), their coefficient of variation (CV) were 38.5%, 36.1% and 46.5% under soil Cd treatments L, M and H, respectively. In treatment L (soil Cd 0.129 mg kg~(-1)), shoot Cd concentrations in all tested cultivars were lower than the ML of Cd allowed by CAC standards (0.2 mg kg~(-1) FW). Thirteen of the tested cultivars measured up to the standard in treatment M (soil Cd 0.352 mg kg~(-1)), 53.6% tested cultivars being higher than the limitation. However, none of the tested cultivars measured up to the standard in treatment treatment H (soil Cd 1.253 mg kg~(-1)). Thus, no Cd-PSC could be found when soil Cd concentration higher than 0.352 mg kg~(-1). only 6 cultivars(No.3、6、7、8、10、11)with lower shoot Cd concentrations and higher shoot biomasses, could be treated as Cd-PSCs, and 3 cultivars(No.19、23、24)with higher shoot Cd concentrations and higher shoot biomasses, could be treated as non-Cd-PSC in the follow-up study.
4. The other pot experiment was carried out to study the variations of accumulation of Cd and Pb by six Cd-PSCs and one non-Cd-PSC. Variations of Cd and Pb concentrations in shoot from genotypes, soil Cd or Pb level and genotype×soil Cd or Pb level were all significant at p < 0.01 level, under lower 0.352 mg kg~(-1)soil Cd level and lower 247 mg kg~(-1)soil Pb level, 4 cultivars(No.7、8、10、11)could be treated as Cd+Pb-PSCs.
5. Detected the seeds Cd concentration in typical Cd accumulation cultivars of celery, lettuce, water spinach, amaranth and Chinese flowering cabbage that screened in early experiments, the results show that, the significant differences were observed between low-Cd Cultivars and high-Cd cultivars of celery, and the seeds Cd concentration of the tested cultivars of celery was significantly correlated with those shoot Cd concentration in pot experiment. So we can use the seed Cd concentration to rapid identify the low-Cd cultivars of celery.
6. Sequential extraction method was used to analyze and distinguish NaCl extractable forms and HCl extractable forms of Cd in seedlings of typical Cd accumulation cultivars of lettuce, water spinach, Chinese cabbage and Chinese flowering cabbage that screened in early experiments, exposed to Cd. The studies showed that, it can be very effective distinguish low-Cd and high-Cd accumulative cultivars for Chinese cabbage and water spinach by using the ratio value of NaCl extractable Cd : HCl extractable Cd. But for the other vegetable species, it should be studied further.
7. In an effort to understand the single nucleotide polymorphisms (SNP) in LdPSC1 gene in 10 typical Cd accumulative cultivars of lettuce, we discovered the specific mutations existed between low-Cd cultivars and high-Cd cultivars in LdPSC1 gene. The test of departure of neutral mutation of nucleotide variation in LdPSC1 gene showed there was a tendency to divide in low-Cd cultivars and high-Cd cultivars, the LdPSC1 gene experiencing strong negative selection under Cd stress. The results could provide a theoretical basis for using the molecular markers in breeding of low-Cd cultivars.
引文
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