6个山黧豆品种的β-ODAP含量与蛋白酶抑制剂活性及其关系分析

Analysis of the Possible Relationship between β-ODAP Content and Protease Inhibitor Activities in Six Lathyrus spp. Cultivars

山黧豆(Lathyrus spp.)种质资源评价与筛选长期聚焦于获得低β-N-草酰-L-α,β-二氨基丙酸(β-N-oxalyl-L-α,β-diaminopropionicacid,β-ODAP)含量或低蛋白酶抑制剂活性的品种。为了研究β-ODAP含量与蛋白酶抑制剂活性之间的潜在关系,该研究利用SDS-PAGE、酶活检测和相关性分析等方法对家山黧豆(L.sativus)、扁荚山黧豆(L.cicera)等6个山黧豆品种的蛋白酶抑制剂活性、β-ODAP含量及二者之间的关系进行了分析。结果表明:(1)不同山黧豆品种的醇溶蛋白和可溶蛋白电泳图谱呈现出显著的差异。(2)不同山黧豆种子的胰蛋白酶抑制剂活性介于100~190 TIU之间,胰凝乳蛋白酶抑制剂活性介于5~9 CIU之间。(3)β-ODAP含量与丝氨酸乙酰基转移酶活性的皮尔逊相关系数可达0.76,呈显著正相关关系。(4)山黧豆转录组数据(SRP145030)中的β-ODAP生物合成关键基因与Bowman-Birk蛋白酶抑制剂(Bowman-Birk inhibitor, BBi)基因表达水平呈显著负相关关系。该研究结果为进一步探讨分析山黧豆β-ODAP和BBi生物合成的调控与平衡,改善山黧豆的含硫氨基酸水平等奠定了基础。

For long time it lasted of evaluation and screening of germplasm resources from different Lathyrus cultivars to get novel lines with low β-N-oxalyl-L-α, β-diaminopropionicacid(β-ODAP) content or protease inhibitor activities. To investigate the possible relationship between β-ODAP content and protease inhibitor activities, we conducted several tests including sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE), enzymatic measurement, correlation analysis etc. referring to six cultivars with two closely related species L. sativus and L. cicera included. The results suggested that:(1) SDS-PAGE analysis of seed globulins and prolamins revealed significant difference in their protein profiles.(2) Kunitz trypsin inhibitor(KTi) activity in different Lathyrus cultivars ranged between 100 and 190 trypsin inhibitor unit, while Bowman-Birk inhibitor(BBi) activity ranged between 5 and 9 chymotrypsin inhibitor unit.(3) The Pearson’s correlation coefficient up to 0.76 suggested a significant positive correlation between β-ODAP content and serine acetyltransferase activity.(4) Correlation analysis of β-ODAP biosynthesis related genes and BBi genes expression from transcriptomic database(SRP145030) suggested a significant negative correlation between them. These results will help to further investigate the regulation of β-ODAP and BBi biosynthesis and improve sulfur amino acid content in Lathyrus cultivars.