RESUMO
Virus-induced gene silencing (VIGS) technology was applied to silence VvANR in cv. Zaoheibao grape berries, and the effects of VvANR silencing on berries phenotype; gene expression level of ANS, LAR1, LAR2, and UFGT; enzyme activity of ANS; and accumulations of anthocyanin and flavan-3-ol were investigated. At the third day after treatment, the VvANR silenced grape berries began to turn red slightly, which was 2 days earlier than that of the control group. And the flavan-3-ol content in VvANR-silenced grape berries had been remarkable within 1 to 5 days, the ANR enzyme activity in VvANR-silenced grapes extremely significantly decreased in 3 days, and LAR enzyme activity also decreased, but the difference was not striking. The ANS enzyme activity of the transformed berries was significantly higher than that of the control after 3 days of infection, and it was exceedingly significantly higher than that of the control after 5 to 10 days. The content of anthocyanin in transformed berries increased of a very marked difference within 3 to 15 days. pTRV2-ANR infection resulted in an extremely significant decrease in the expression of VvANR gene, and the expression of VvLAR1, VvLAR2, VvMYBPA1, VvMYBPA2, and VvDFR were also down-regulated. However, the expression of VvANS and VvUFGT was up-regulated significantly. After VvANR silencing via VIGS, VvANR expression in grape berries was extremely significantly decreased, resulting in decreased ANR enzyme activity and flavan-3-ol content; berries turned red and deeper in advance. In addition, VvANR silencing can induce up-regulation of VvANS and VvUFGT expression, significantly increase ANS enzyme activity, and increase of anthocyanin accumulation.
Assuntos
Vitis , Antocianinas/metabolismo , Frutas/genética , Frutas/metabolismo , Regulação da Expressão Gênica de Plantas , Ativação Transcricional , Vitis/metabolismoRESUMO
Glycinebetaine (GB) is an osmoprotectant found in plants under environmental stresses that incorporates drought and is associated with drought tolerance in several plants, such as the woody pear. However, how GB improves drought tolerance in pears remains unclear. In the current study, we explored the mechanism by which GB enhances drought tolerance of whole pear plants (Pyrus bretschneideri Redh. cv. Suli) supplied with exogenous GB. The results showed that on the sixth day after withholding water, levels of O2·-, H2O2, malonaldehyde (MDA) and electrolyte leakage in the leaves were substantially increased by 143%, 38%, 134% and 155%, respectively. Exogenous GB treatment was substantially reduced O2·-, H2O2, MDA and electrolyte leakage (38%, 24%, 38% and 36%, respectively) in drought-stressed leaves. Furthermore, exogenous GB induced considerably higher antioxidant enzyme activity in dry-stressed leaves than drought-stressed treatment alone on the sixth day after withholding water, such as superoxide dismutase (SOD) (201%) and peroxidase (POD) (127%). In addition, these GB-induced phenomena led to increased endogenous GB levels in the leaves of the GB 100 + drought and GB 500 + drought treatment groups by 30% and 78%, respectively, compared to drought treatment alone. The findings obtained were confirmed by the results of the disconnected leaf tests, in which GB contributed to a substantial increase in SOD activity and parallel dose- and time-based decreases in MDA levels. These results demonstrate that GB-conferred drought resistance in pears may be due in part to minimizing symptoms of oxidative harm incurred in response to drought by the activities of antioxidants and by reducing the build-up of ROS and lipid peroxidation.