青花菜中莱菔硫烷调控基因QTL精细定位

Sulforaphane is one of the strongest ingredients of anti-cancer in all of the vegetables. Broccoli (Brassica oleracea L. var. italica) is the highest vegetables in containing sulforaphane. The current study showed that the synthesis of sulforaphane is mainly regulated by genes, and the generation of sulforaphane depends on its precursor glucoraphanin. In the process of glucoraphanin accumulation, multiple family genes are involved. So far, it is unclear how many genes regulate the generation of sulforaphane in the metabolism of glucosinolate. In the study, one F1DH population derived from two inbred line of broccoli lines will be used by isolated microspore culture, and there was a significant difference in the two inbred lines of broccoli. At the same time, more than 8 000 markers including SSR, InDel and SNP will be designed for construction of a high density genetic linkage map. In the study, sulforaphane content will be detected by HPLC. Finally the software MapQTL 4.0 will be used for QTL mapping of sulforaphane content by the F1DH population. The main QTL will be located in the Brassica genome by the annotation genomes of cabbage, Chinese cabbage and Arabidopsis. The study will lay a foundation for cloning of sulforaphane regulated genes, meanwhile materials with higher content of sulforaphane and new varieties of broccoli and the other Brassica vegetables would be promoted by molecular marker assistant.

青花菜富含莱菔硫烷抗癌活性成分，莱菔硫烷的合成主要受基因型调控，而莱菔硫烷的生成量主要取决于其前体4-甲基亚磺酰基丁基硫甙的合成，4-甲基亚磺酰基丁基硫甙的合成代谢过程中涉及多个基因家族。目前尚不清楚青花菜中莱菔硫烷含量是由哪些硫甙基因调控。本研究拟以花球中莱菔硫烷含量差异显著的2份青花菜高代纯合自交系配制的F1，经小孢子培养获得的DH群体为试验材料，利用8000多对SSR、InDel和SNP标记构建青花菜高密度遗传连锁图谱。利用HPLC技术对青花菜DH群体花球中莱菔硫烷含量进行检测，采用MapQTL 4.0软件对青花菜DH群体花球中莱菔硫烷含量进行QTL精细定位，通过已注释的甘蓝、白菜和拟南芥基因组将主效QTL在甘蓝基因组上进行锚定。该研究为莱菔硫烷调控基因的克隆奠定基础，有利于选育出莱菔硫烷含量较高的青花菜材料和甘蓝类蔬菜新品种。

青花菜富含莱菔硫烷抗癌活性成分，莱菔硫烷的合成主要受基因型调控，而莱菔硫烷的生成量主要取决于其前体4-甲基亚磺酰基丁基硫甙的合成，4-甲基亚磺酰基丁基硫甙的合成代谢过程中涉及多个基因家族。目前尚不清楚青花菜中莱菔硫烷含量是由哪些硫甙基因调控。本研究拟以花球中莱菔硫烷含量差异显著的2份青花菜高代纯合自交系配制的F1，经小孢子培养获得的DH群体为试验材料，利用8042对SSR、InDel和SNP引物最终构建了一个包含9条连锁群，包含268对分子标记的遗传连锁图谱，该图谱覆盖基因组长度1293 cM，最小遗传距离达到了1.68 cM。利用HPLC技术对青花菜DH群体花球中莱菔硫烷含量进行了分析，采用MapQTL 4.0软件对青花菜DH群体花球中莱菔硫烷含量进行QTL精细定位，获得13个相关QTL，在SF-1QTL区间加密，获得了一个候选基因BOSF（Bol039589）。该研究为莱菔硫烷调控基因的克隆奠定了基础，为青花菜重要农艺性状的定位和基础研究提供了前提。