Over-expression of a fungal NADP(H)-dependent glutamate dehydrogenase PcGDH improves nitrogen assimilation and growth quality in rice

Zhou, Yunhua; Liu, Hong; Zhou, Xiangcheng; Yan, Yuanzhu; Du, Changqing; Li, Yixing; Liu, Derong; Zhang, Caisheng; Deng, Xiaolong; Tang, Dongying; Zhao, Xiaoying; Zhu, Yonghua; Lin, Juan; Liu, Xuanming

doi:10.1007/s11032-014-0037-z

Cited by 31 publications

(41 citation statements)

References 52 publications

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“…1d), indicating that the capacity for ammonium assimilation of His-TF-MgGDH was higher than that of His-TF-OsGDH. Furthermore, the K m value of MgGDH (0.67 ± 0.05 mM) is lower than those of previous reported fungal gdhA (1.16 ± 0.13 mM; Abiko et al 2010) and PcGDH (3.73 ± 0.23 mM; Zhou et al 2014b), previously shown to improve ammonium assimilation in transgenic rice, indicating that MgGDH has a higher capacity to assimilate ammonium than gdhA and PcGDH.…”

Section: Resultscontrasting

confidence: 70%

“…2d), which indicated that MgGDH is a cytoplasm-localized protein. In agreement with this observation, it was recently reported that fungal GDH protein gdhA (Abiko et al 2010), CeGDH (Zhou et al 2014a) and PcGDH (Zhou et al 2014b) were also cytoplasm-localized.…”

Section: Characteristics Of Transgenic Rice Plants Over-expressing Mggdhsupporting

confidence: 78%

“…In contrast, the affinity of NADP(H)-GDH from microorganism for ammonium is much higher (Wang and Tian 2001;Noor et al 2005;Abiko et al 2010;Du et al 2014;Zhou et al 2014b). For example, the K m value of NiGDH for ammonium has been reported to range from 0.3 to 0.45 mM (Wang and Tian 2001), while that of gdhA ranges from 1.05 mM (Noor et al 2005) to 1.25 mM (Abiko et al 2010).…”

Section: Discussionmentioning

confidence: 99%

“…Both purified His-TF-OsGDH and His-TFMgGDH fusion proteins were assayed for enzymatic activity in vitro. The NADP(H)-GDH activities of His-TFOsGDH and His-TF-MgGDH were determined as described previously (Zhou et al 2014b). One unit of enzyme activity is defined as the reduction or oxidation of 1 lM of NADP(H) per minute per microgram His-TF-OsGDH and His-TF-MgGDH at 25°C.…”

Section: Plant Materials and Stress Treatmentmentioning

confidence: 99%

“…The concentration of glutamate in the shoots and roots of transgenic and wild-type plants was determined using commercial kits (NJJCBIO, Nanjing, China) as described previously (Zhou et al 2014b). Briefly, leaves (approximately 0.1 g FW) were homogenized in 0.9 mL plant protein extraction reagent (CWBIO, Beijing, China) for 20 min on ice.…”

Section: Real-time Pcr Analysismentioning

confidence: 99%

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Over-expression of a glutamate dehydrogenase gene, MgGDH, from Magnaporthe grisea confers tolerance to dehydration stress in transgenic rice

Zhou

Zhang

Lin

et al. 2014

Planta

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Heterologous expression of a fungal NADP(H)-GDH gene ( MgGDH ) from Magnaporthe grisea can improve dehydration stress tolerance in rice by preventing toxic accumulation of ammonium. Glutamate dehydrogenase (GDH; EC 1.4.1.2 and EC 1.4.1.4) may act as a stress-responsive enzyme in detoxification of high intracellular ammonia and production of glutamate for proline synthesis under stress conditions. In present study, a fungal NADP(H)-GDH gene (MgGDH) from Magnaporthe grisea was over-expressed in rice (Oryza sativa L. cv. 'kitaake'), and the transgenic plants showed the improvement of tolerance to dehydration stress. The kinetic analysis showed that His-TF-MgGDH preferentially utilizes ammonium to produce L-glutamate. Moreover, the affinity of His-TF-MgGDH for ammonium was dramatically higher than that of His-TF-OsGDH for ammonium. Over-expressing MgGDH transgenic rice plants showed lower water-loss rate and higher completely close stomata than the wild-type plants under dehydration stress conditions. In transgenic plants, the NADP(H)-GDH activities were markedly higher than those in wild-type plants and the amination activity was significantly higher than the deamination activity. Compared with wild-type plants, the transgenic plants accumulated much less NH4 (+) but higher amounts of glutamate, proline and soluble sugar under dehydration stress conditions. These results indicate that heterologous expression of MgGDH can prevent toxic accumulation of ammonium and in return improve dehydration stress tolerance in rice.

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Section: Resultscontrasting

confidence: 70%

Section: Characteristics Of Transgenic Rice Plants Over-expressing Mggdhsupporting

confidence: 78%