背景:竹子在原处有一个很长的幼年阶段,但可以在体外组织培养过程中诱导开花,为生殖生物学和开花研究提供了现成的材料来源。在这份报告中,通过使用两个测序平台:Illumina和454,收获了来自体外的维德芽孢杆菌的生殖和营养材料,并用于产生转录组数据库。两个数据集的组合导致高转录组质量和增加的序列读段长度。在植物中,许多MADS基因控制花卉发育,已经开发了ABCDE模型来解释基因如何共同作用以在花中创建不同的螺纹。
结果:作为案例研究,已发表的与水稻花发育相关的OsMADS蛋白用于搜索B.edulis转录组数据集,识别16B.edulisMADS(BeMADS)。通过qRT-PCR和原位杂交测定BeMADS基因表达水平。大多数BeMADS基因在花中高表达,除了BeMADS34。这些基因的表达模式与水稻同源物最为相似,除BeMADS18和BeMADS34外,与水稻花发育ABCDE模型高度相似。MADS-GFP蛋白的瞬时表达表明只有BeMADS1进入叶核。BeMADS18、BeMADS4和BeMADS1位于外核。当与BeMADS1共转化时,BeMADS15、16、13、21、6和7在引理中易位到细胞核,表明BeMADS1是其他BeMADS亚细胞定位的关键因素。
结论:我们的研究提供了丰富的B.edulis转录组数据,并提供了全面的序列资源。结果,此处报道的分子材料和总体策略可用于未来的基因鉴定和对竹子经济上重要的作物的进一步生殖研究。
BACKGROUND: The bamboo Bambusa edulis has a long juvenile phase in situ, but can be induced to flower during in vitro tissue culture, providing a readily available source of material for studies on reproductive biology and flowering. In this report, in vitro-derived reproductive and vegetative materials of B. edulis were harvested and used to generate transcriptome databases by use of two sequencing platforms: Illumina and 454. Combination of the two datasets resulted in high transcriptome quality and increased length of the sequence reads. In plants, many MADS genes control flower development, and the ABCDE model has been developed to explain how the genes function together to create the different whorls within a flower.
RESULTS: As a case
study, published floral development-related OsMADS proteins from rice were used to search the B. edulis transcriptome datasets, identifying 16 B. edulis MADS (BeMADS). The BeMADS gene expression levels were determined qRT-PCR and in situ hybridization. Most BeMADS genes were highly expressed in flowers, with the exception of BeMADS34. The expression patterns of these genes were most similar to the rice homologs, except BeMADS18 and BeMADS34, and were highly similar to the floral development ABCDE model in rice. Transient expression of MADS-GFP proteins showed that only BeMADS1 entered leaf nucleus. BeMADS18, BeMADS4, and BeMADS1 were located in the lemma nucleus. When co-transformed with BeMADS1, BeMADS15, 16, 13, 21, 6, and 7 translocated to nucleus in lemmas, indicating that BeMADS1 is a key factor for subcellular localization of other BeMADS.
CONCLUSIONS: Our
study provides abundant B. edulis transcriptome data and offers comprehensive sequence resources. The results, molecular materials and overall strategy reported here can be used for future gene identification and for further reproductive studies in the economically important crop of bamboo.