International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Patancheru, Hyderabad 502 324, India.
International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Patancheru, Hyderabad 502 324, India.
Int J Biol Macromol. 2020 Aug 1;156:103-110. doi: 10.1016/j.ijbiomac.2020.04.069. Epub 2020 Apr 12.
In the present study, the promoter region of the pearl millet heat shock protein 10 (PgHsp10) gene was cloned and characterized. The PgHsp10 promoter (PgHsp10pro) sequence region has all the cis-motifs required for tissue and abiotic stress inducibility. The complete PgHsp10pro (PgHsp10PC) region and a series of 5' truncations of PgHsp10 (PgHsp10D1 and PgHsp10D2) and an antisense form of PgHsp10pro (PgHsp10AS) were cloned into a plant expression vector (pMDC164) through gateway cloning. All four constructs were separately transformed into tobacco through Agrobacterium-mediated genetic transformation, and PCR-confirmed transgenic plants progressed to T and T generations. The T transgenic tobacco plants comprising all PgHsp10pro fragments were used for GUS histochemical and qRT-PCR assays in different tissues under control and abiotic stresses. The PgHsp10PC pro expression was specific to stem and seedlings under control conditions. Under different abiotic stresses, particularly heat stress, PgHsp10PCpro had relatively higher activity than PgHsp10D1pro, PgHsp10D2pro and PgHsp10ASpro. PgHsp10pro from a stress resilient crop like pearl millet responds positively to a range of abiotic stresses, in particular heat, when expressed in heterologous plant systems such as tobacco. Hence, PgHsp10pro appears to be a potential promoter candidate for developing heat and drought stress-tolerant crop plants.
在本研究中,克隆并鉴定了珍珠粟热休克蛋白 10(PgHsp10)基因的启动子区域。PgHsp10 启动子(PgHsp10pro)序列区域具有组织和非生物胁迫诱导所需的所有顺式基序。完整的 PgHsp10pro(PgHsp10PC)区域和一系列 5'截短的 PgHsp10(PgHsp10D1 和 PgHsp10D2)以及 PgHsp10pro 的反义形式(PgHsp10AS)通过 gateway 克隆被克隆到植物表达载体(pMDC164)中。这四个构建体分别通过农杆菌介导的遗传转化转化到烟草中,并通过 PCR 确认转化的植物进入 T 和 T 代。包含所有 PgHsp10pro 片段的 T 代转基因烟草植物在不同组织中进行 GUS 组织化学和 qRT-PCR 分析,在对照和非生物胁迫下。在对照条件下,PgHsp10PCpro 特异性表达于茎和幼苗。在不同的非生物胁迫下,特别是热胁迫下,PgHsp10PCpro 的活性相对高于 PgHsp10D1pro、PgHsp10D2pro 和 PgHsp10ASpro。在烟草等异源植物系统中表达时,来自像珍珠粟这样具有抗逆性的作物的 PgHsp10pro 对一系列非生物胁迫,特别是热胁迫,有积极的响应。因此,PgHsp10pro 似乎是开发耐热和耐旱作物的潜在启动子候选者。