Transcriptomics Lab (K-Lab), Division of Plant Biotechnology, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Srinagar, J&K, 190025, India.
Division of Vegetable Science, SKUAST-K, Srinagar, J&K, 190025, India.
Mol Biol Rep. 2022 Jun;49(6):5229-5240. doi: 10.1007/s11033-021-06619-z. Epub 2021 Aug 13.
Sea buckthorn (Hippophae) is in the focus of interest mainly for its positive effects on health of both human and animal organisms. Due to the similarities in vegetative morphology, Hippophae species are often misidentified. Therefore, current study was focused on ITS based sequence characterization of sea buckthorn species and comparative biochemical evaluation for its antioxidant properties.
DNA was extracted from leaf samples. Primer pairs K-Lab-SeaBukRhm-ITS1F1- K-Lab-SeaBukRhm-ITS1R1 and K-LabSeaBukTib- ITSF1- K-LabSeaBukTib-ITSR1 were used for PCR amplification. The purified PCR products were outsourced for sequencing. Phylogenetic tree was constructed based on neighbor-joining (NJ) method. Moreover, comparison of antioxidant potential of leaves of two sea buckthorn species (Hippophae rhamnoides and Hippophae tibetana) collected from different regions of Ladakh viz., Stakna, Nubra, DRDO Leh and Zanskar was determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-bis (3- ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS), and Total antioxidant capacity (TAC) by phosphomolybdenum assays. The present investigation led to the differentiation of two sea buckthorn species viz., H. rhamnoides and H. tibetana based on Internal Transcribed Spacer (ITS) region. Moreover, significant variation was observed in antioxidant potential of leaf extracts collected from different regions.
Primary ITS sequence analysis was found to be powerful tool for identification and genetic diversity studies in sea buckthorn. Leaves of sea buckthorn have pronounced antioxidant properties and can be used in food, neutraceuticals and pharmaceutical industries etc. The current study will pave the way to discover small bioactive molecules responsible for antioxidant and anticancer properties in sea buckthorn.
沙棘（Hippophae）主要因其对人类和动物生物体健康的积极影响而受到关注。由于在营养形态上的相似性，沙棘物种经常被错误识别。因此，本研究主要集中在ITS 序列特征分析沙棘物种和比较其抗氧化特性的生化评估上。
从叶片样本中提取 DNA。使用引物对 K-Lab-SeaBukRhm-ITS1F1-K-Lab-SeaBukRhm-ITS1R1 和 K-LabSeaBukTib-ITSF1-K-LabSeaBukTib-ITSR1 进行 PCR 扩增。纯化的 PCR 产物外包测序。基于邻接法（NJ）构建系统发育树。此外，还通过 2,2-二苯基-1-苦基肼基（DPPH）、2,2-联氮-双（3-乙基苯并噻唑啉-6-磺酸）二铵盐（ABTS）和磷钼酸盐法测定的总抗氧化能力（TAC）来确定来自拉达克不同地区（Stakna、Nubra、DRDO 列城和赞斯卡）的两种沙棘（Hippophae rhamnoides 和 Hippophae tibetana）叶片的抗氧化潜力。本研究通过内转录间隔区（ITS）区将两种沙棘物种区分开来。此外，还观察到来自不同地区的叶片提取物的抗氧化潜力存在显著差异。
初步的 ITS 序列分析被发现是沙棘鉴定和遗传多样性研究的有力工具。沙棘叶具有明显的抗氧化特性，可用于食品、营养保健品和制药等行业。本研究将为发现负责沙棘抗氧化和抗癌特性的小分子生物活性物质铺平道路。
