Mall Maneesha, Shanker Karuna, Samad Abdul, Kalra Alok, Sundaresan Velusamy, Shukla Ashutosh K
CSIR-Central Institute of Medicinal and Aromatic Plants, P.O. CIMAP, Lucknow, 226015, UP, India.
CSIR-Central Institute of Medicinal and Aromatic Plants, Research Centre, Allalasandra, GKVK Post, Bengaluru, 560065, Karnataka, India.
Protoplasma. 2022 May;259(3):755-773. doi: 10.1007/s00709-021-01701-6. Epub 2021 Aug 30.
Vindoline is an important alkaloid produced in Catharanthus roseus leaves. It is the more important monomer of the scarce and costly anticancer bisindole alkaloids, vincristine, and vinblastine, as unlike catharanthine (the other monomer), its biosynthesis is restricted to the leaves. Here, biotic (bacterial endophyte, phytoplasma, virus) and abiotic (temperature, salinity, SA, MeJa) factors were studied for their effect on vindoline accumulation in C. roseus. Variations in vindoline pathway-related gene expression were reflected in changes in vindoline content. Since allene oxide cyclase (CrAOC) is involved in jasmonate biosynthesis and MeJa modulates many vindoline pathway genes, the correlation between CrAOC expression and vindoline content was studied. It was taken up for full-length cloning, tissue-specific expression profiling, in silico analyses, and upstream genomic region analysis for cis-regulatory elements. Co-expression analysis of CrAOC with vindoline metabolism-related genes under the influence of aforementioned abiotic/biotic factors indicated its stronger direct correlation with the tabersonine-to-vindoline genes (t16h, omt, t3o, t3r, nmt, d4h, dat) as compared to the pre-tabersonine genes (tdc, str, sgd). Its expression was inversely related to that of downstream-acting peroxidase (prx) (except under temperature stress). Direct/positive relationship of CrAOC expression with vindoline content established it as a key gene modulating vindoline accumulation in C. roseus.
长春多灵是长春花叶片中产生的一种重要生物碱。它是稀缺且昂贵的抗癌双吲哚生物碱长春新碱和长春碱中更重要的单体,与长春质碱(另一种单体)不同,其生物合成仅限于叶片。在此,研究了生物因素(细菌内生菌、植原体、病毒)和非生物因素(温度、盐度、水杨酸、茉莉酸甲酯)对长春花中长春多灵积累的影响。长春多灵途径相关基因表达的变化反映在长春多灵含量的变化上。由于丙二烯氧化物环化酶(CrAOC)参与茉莉酸生物合成,且茉莉酸甲酯调节许多长春多灵途径基因,因此研究了CrAOC表达与长春多灵含量之间的相关性。对其进行了全长克隆、组织特异性表达谱分析、计算机分析以及上游基因组区域顺式调控元件分析。在上述非生物/生物因素影响下,对CrAOC与长春多灵代谢相关基因进行共表达分析,结果表明,与长春多灵生物合成前体长春质碱相关基因(tdc、str、sgd)相比,它与从利血平宁到长春多灵的基因(t16h、omt、t3o、t3r、nmt、d4h、dat)的直接相关性更强。其表达与下游作用的过氧化物酶(prx)的表达呈负相关(温度胁迫下除外)。CrAOC表达与长春多灵含量的直接/正相关关系表明它是调节长春花中长春多灵积累的关键基因。