当前位置: X-MOL 学术Gene › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Relative expression of putative genes involved in galanthamine and other Amaryllidaceae alkaloids biosynthesis in Narcissus field and in vitro tissues
Gene ( IF 2.6 ) Pub Date : 2021-01-09 , DOI: 10.1016/j.gene.2021.145424
Ferdausi Aleya , Chang Xianmin , Hall Anthony , Jones Meriel

The Narcissus pseudonarcissus cv. Carlton contains Amaryllidaceae alkaloids namely galanthamine, lycorine, homolycorine, narciclasine, which are noted for their pharmaceutical properties such as for the treatment of early to mid-stage Alzheimer's diseases, cancer, tumor etc. Alkaloid biosynthesis using plant in vitro systems has been considered as a tool for drug discovery and the pathways are starting to be understood but still far from complete. Therefore, the study was emphasized to observe the relative expressions of putative genes involved in the biosynthetic pathway leading to the Amaryllidaceae alkaloids in field grown bulbs and developing cell culture systems in Narcissus.

MS media fortified with growth regulators were used for the development of tissue culture from Carlton twin-scale explants. MS medium with high auxin, 20 mg/l NAA was the best medium for callus growth and maintenance while media with low auxin, 4 mg/l NAA and MS basal media gave the maximum bulblets. Field tissues showed a higher amount of galanthamine content; i.e. basal plate (1050–1310 µg Gal/g FW) and bulb (980–1150 µg Gal/g FW) than the culture derived samples; callus (1.0–7.0 µg Gal/g FW) and bulblets (12–215 µg Gal/g FW) on a fresh weight (FW) basis. GC–MS chromatograms of samples under study also showed the presence of other important alkaloids i.e. lycorine, homolycorine, lycorenine, haemanthamine, crinamine, lycoramine and tazettine. RNA extracted from in vitro callus, bulblets and field grown bulb, basal plate were used for PCR to detect the relative expression of putative genes; P450, PAL, TYDC and NpO4OMT normalized to actin. The selected transcripts for P450s and TYDC were expressed in both field and in vitro tissues. Higher expressions of PAL were observed in calli than field samples. The expression of NpN4OMT was notably higher in field samples than in vitro tissues. Therefore, in vitro tissues could be a good source for the reproducible and easy extraction of alkaloids from plants.



中文翻译:

水仙花场和体外组织中涉及加兰他敏和其他金莲花科生物碱合成的推定基因的相对表达

水仙品种。卡尔顿包含石蒜科生物碱即加兰他敏,石蒜碱,homolycorine,水仙环素,其为它们的药学特性,如治疗的初期到中期阶段阿尔茨海默氏病,癌症,肿瘤注意到等生物碱合成使用植物体外系统已被认为是一种药物发现的工具,其途径已开始被理解,但仍远未完成。因此,强调该研究的目的是观察田间生长的鳞茎和发育中的水仙的生物合成途径中导致金缕梅生物碱的生物合成途径的推定基因的相对表达。

用生长调节剂强化的MS培养基用于从Carlton双尺度外植体开发组织培养物。高生长素,20 mg / l NAA的MS培养基是愈伤组织生长和维持的最佳培养基,而低生长素,4 mg / l NAA的培养基和MS基础培养基的球茎最大。野外组织显示出更高的加兰他敏含量。例如,基底板(1050-1310 µg Gal / g FW)和球茎(980-1150 µg Gal / g FW)要比培养样品高;以鲜重(FW)为基础的愈伤组织(1.0–7.0 µg Gal / g FW)和鳞茎(12–215 µg Gal / g FW)。所研究样品的GC-MS色谱图还显示了其他重要生物碱的存在,例如,茄碱,高茄碱,lycorenine,山梨糖胺,克林胺,lycoramine和他塞汀。体外提取RNA用愈伤组织,鳞茎和田间生长的鳞茎,基底板进行PCR,以检测推定基因的相对表达。P450,PALTYDCNpO 4 OMT标准化为肌动蛋白。P450和TYDC的选定转录本在野外和体外组织中均表达。与现场样品相比,在愈伤组织中观察到PAL的表达更高。Np N4OMT在野外样品中的表达明显高于体外组织。因此,体外组织可能是从植物中可再生且容易提取生物碱的良好来源。

更新日期:2021-01-22
down
wechat
bug