花生耐盐乙烯不敏感突变体的筛选

Screening of Peanut Mutants with Salt Tolerance and Ethylene Insensitivity

利用沙土培养法,使用1、3、5、7、10 mg·L^-1乙烯利溶液处理花生品种花育22种子,5 d后测量下胚轴长度。结果表明:经乙烯利处理后,花生下胚轴生长受到显著抑制而增粗变短,受抑制程度随乙烯利溶液浓度的提高而增加,3 mg·L^-1乙烯利处理的下胚轴长度极显著低于未施加和施加1 mg·L^-1乙烯利处理的,因此选用3 mg·L^-1乙烯利对诱变筛选得到的165份花生种质进行种子萌发处理,有145份种质表现为下胚轴生长受抑制而增粗变短,属于正常的乙烯敏感型种质;有11份种质下胚轴生长并未受到明显抑制,显著高于同样条件处理下花育22的下胚轴长度,属于乙烯不敏感型种质;有9份种质下胚轴生长受到显著抑制,其下胚轴长度极显著低于同样条件处理下花育22的下胚轴长度,属于乙烯超敏感型种质。使用50μmol·L^-1 ACC(1-氨基环丙烷羧酸)溶液对筛选获得的11份不敏感种质进行验证处理,其中,9份种质保持了对乙烯不敏感的特点,下胚轴生长仍未受到明显抑制。通过在沙土中添加0.7%NaCl溶液,继续对筛选得到的9份乙烯不敏感型种质进行耐盐筛选,有1份种质表现为下胚轴生长受抑制程度最低,下胚轴长度显著高于同样处理条件下花育22的下胚轴长度,初步确定其为耐盐乙烯不敏感种质。因此,用3 mg·L^-1乙烯利溶液预选、50μmol·L^-1 ACC溶液验证、0.7%NaCl溶液处理,通过评价黑暗条件下沙土培养的花生种子下胚轴长度,可以有效筛选到花生耐盐乙烯不敏感突变体。

Seeds of peanut cultivar Huayu 22 were cultured in sands with 1, 3, 5, 7 and 10 mg·L^-1 ethephon solution, and the hypocotyl length was measured after five days. The results showed that the hypocotyl growth was significantly inhibited and got shorter and weaker after treated with ethephon, and the inhibition increased with the increasing concentration of ethephon solution. Hypocotyl length treated with 3 mg·L^-1 ethephon was significantly shorter than that treated with 1 mg·L^-1 ethephon or not,So one hundred and sixty-five peanut germplasm accessions from mutagenesis were treated and screened with 3 mg·L^-1 ethephon solution. The obvious hypocotyl growth inhibition was found in 145 accessions which belonged to the ethylene sensitive accessions. The hypocotyl growth of 11 accessions were not inhibited obviously, and were significantly higher than that of Huayu 22 under the same treatment, which belonged to the ethylene insensitive accessions. The significantly obvious hypocotyl growth inhibition was found in 9 accessions with the hypocotyl length significantly lower than that of Huayu 22 under the same treatment, which belonged to the ethylene super sensitive accessions. The seeds of selfed progenies from 11 insensitive accessions were cultured in sands with 50 μmol·L^-1 ACC solution, and there were 9 accessions whose hypocotyl growth was still not inhibited obviously. Salt tolerance screening was conducted by adding 0.7% NaCl solution in sands for the 9 stable insensitive accessions. There was one accession whose hypocotyl growth was obviously higher than that of Huayu 22 under the same treatment, which was regarded as a salt tolerant and ethylene insensitive accession. Therefore, the salt tolerant and ethylene insensitive mutants of peanut could be screened by pre-selection with 3 mg·L^-1 ethephon solution, validation with 50 μmol·L^-1 ACC solution,treating with 0.7% NaCl solution and evaluating the hypocotyl length of peanut seeds cultured in sandy soil under dark conditions.