Molecular Cloning and Expression Analysis of the ζ-Carotene Desaturase Gene in Chinese kale(Brassica oleracea var.alboglabra Bailey)

ζ-Carotene desaturase(ZDS)is an important enzyme in carotenoid biosynthesis.Here,the Brassica oleracea var.alboglabra ZDS(Boa ZDS)gene was cloned from Chinese kale via reverse transcription-polymerase chain reaction(RT-PCR)and deposited in Gen Bank(accession number KY662297).The Boa ZDS gene contains an open reading frame of 1 686 bp that encodes a 561-amino acid protein.Sequence analysis indicates that the ZDS protein is apparently conserved during plant evolution and is most closely related to B.oleracea var.capitata and B.rapa.The promoter sequence of the Boa ZDS gene was predicted to harbor several cis-acting elements that are related to light and phytohormone responses.Semiquantitative RT-PCR analysis showed that Boa ZDS expression varied among different developmental stages and organs.Relative ZDS expression remained stable during germination and seedling stages and rapidly increased at the mature leaf stage.The leaves showed the highest ZDS expression levels compared to the other organs.ZDS expression decreased in all flower tissues during blooming.The fused protein of Boa ZDS was obtained by prokaryotic expression.Heterologous expression of Boa ZDS in Escherichia coli confirmed that Boa ZDS encodes a functionalζ-carotene desaturase that increases β-carotene accumulation in E.coli cells harboring a β-carotene-producing plasmid.The findings of the present study provide a molecular basis for the elucidation of ZDS gene function in Chinese kale.

Germplasm Diversity of Chinese Kale in China

Chinese kale is an important vegetable crop in China, especially in South China where the majority of varieties are grown. It originated in South China and spread throughout Southeast Asia thereafter. Chinese kale can be classified into two types according to whether the petals are white or yellow. There are also three main cultivated types based on the edible organs, including the stalk and leaf type, the stalk type,and the leaf type. Two primary types have also been defined based on stalk color, including green stalks and red stalks. They are also cultivated based on the type of stalk, including main stalk and lateral stalk types. Significant differences have also been observed in glucosinolate content among the varieties, and a 40-fold difference in neoindle-3-methyl glucosinolate was detected across the cultivars.