Li Zhanjie, Cheng Yufeng, Cui Jianmin, Zhang Peipei, Zhao Huixian, Hu Shengwu
State Key Laboratory of Crop Stress Biology in Arid Areas, Northwest A&F University, Yangling, Shaanxi, 712100, P. R. China.
College of Life Sciences, Northwest A&F University, Yangling, Shaanxi, 712100, P.R. China.
BMC Genomics. 2015 Mar 17;16(1):206. doi: 10.1186/s12864-015-1388-5.
Chemical hybridization agents (CHAs) are often used to induce male sterility for the production of hybrid seeds. We previously discovered that monosulfuron ester sodium (MES), an acetolactate synthase (ALS) inhibitor of the herbicide sulfonylurea family, can induce rapeseed (Brassica napus L.) male sterility at approximately 1% concentration required for its herbicidal activity. To find some clues to the mechanism of MES inducing male sterility, the ultrastructural cytology observations, comparative transcriptome analysis, and physiological analysis on carbohydrate content were carried out in leaves and anthers at different developmental stages between the MES-treated and mock-treated rapeseed plants.
Cytological analysis revealed that the plastid ultrastructure was abnormal in pollen mother cells and tapetal cells in male sterility anthers induced by MES treatment, with less material accumulation in it. However, starch granules were observed in chloroplastids of the epidermis cells in male sterility anthers. Comparative transcriptome analysis identified 1501 differentially expressed transcripts (DETs) in leaves and anthers at different developmental stages, most of these DETs being localized in plastid and mitochondrion. Transcripts involved in metabolism, especially in carbohydrate and lipid metabolism, and cellular transport were differentially expressed. Pathway visualization showed that the tightly regulated gene network for metabolism was reprogrammed to respond to MES treatment. The results of cytological observation and transcriptome analysis in the MES-treated rapeseed plants were mirrored by carbohydrate content analysis. MES treatment led to decrease in soluble sugars content in leaves and early stage buds, but increase in soluble sugars content and decrease in starch content in middle stage buds.
Our integrative results suggested that carbohydrate and lipid metabolism were influenced by CHA-MES treatment during rapeseed anther development, which might responsible for low concentration MES specifically inducing male sterility. A simple action model of CHA-MES inducing male sterility in B. napus was proposed. These results will help us to understand the mechanism of MES inducing male sterility at low concentration, and might provide some potential targets for developing new male sterility inducing CHAs and for genetic manipulation in rapeseed breeding.
化学杂交剂(CHAs)常用于诱导雄性不育以生产杂交种子。我们之前发现,单嘧磺酯钠(MES)是除草剂磺酰脲家族的乙酰乳酸合成酶(ALS)抑制剂,在其除草活性所需的约1%浓度下可诱导油菜(甘蓝型油菜L.)雄性不育。为了寻找MES诱导雄性不育机制的线索,对MES处理和模拟处理的油菜植株不同发育阶段的叶片和花药进行了超微结构细胞学观察、比较转录组分析以及碳水化合物含量的生理分析。
细胞学分析表明,MES处理诱导的雄性不育花药中的花粉母细胞和绒毡层细胞的质体超微结构异常,其中物质积累较少。然而,在雄性不育花药的表皮细胞质体中观察到淀粉粒。比较转录组分析确定了不同发育阶段叶片和花药中的1501个差异表达转录本(DETs),这些DETs大多定位于质体和线粒体。参与代谢,尤其是碳水化合物和脂质代谢以及细胞运输的转录本差异表达。通路可视化显示,紧密调控的代谢基因网络被重新编程以响应MES处理。MES处理的油菜植株的细胞学观察和转录组分析结果在碳水化合物含量分析中得到了反映。MES处理导致叶片和早期花蕾中可溶性糖含量降低,但中期花蕾中可溶性糖含量增加而淀粉含量降低。
我们的综合结果表明,在油菜花药发育过程中,碳水化合物和脂质代谢受CHA-MES处理的影响,这可能是低浓度MES特异性诱导雄性不育的原因。提出了CHA-MES诱导甘蓝型油菜雄性不育的简单作用模型。这些结果将有助于我们理解MES在低浓度下诱导雄性不育的机制,并可能为开发新的诱导雄性不育的CHAs以及油菜育种中的基因操作提供一些潜在靶点。