Induced extracellular production of viniferins in grapevine cell culture medium by elicitation with methyl jasmonate and stevioside

We report the high production of stilbenes, including resveratrol and viniferins, in grapevine (Vitis labruscana L.) cell cultures through elicitation with methyl jasmonate (MJ) and stevioside (STE). Methyl-β-cyclodextrin (CD-M) is widely used as a solubilizer for resveratrol production. For the first time, we used STE as a solubilizer for stilbene production in plant cell cultures. MJ was most effective in activating VvSTS expression and stimulating stilbene biosynthesis and induced accumulation of trans-piceid in the cells. High levels of trans-resveratrol and δ-viniferin production were observed in the culture medium when CD-M and STE, respectively, were used as solubilizers, but neither were significantly elevated within the cells. Maximum production of trans-resveratrol (12.2 mg/L) and δ-viniferin (892.2 mg/L) was observed 5 days after elicitation of cells with MJ and STE in shake flask cultures. Notably, predominant production of δ-viniferin and trans-resveratrol was observed in shake and static flask culture medium, respectively. Furthermore, stilbene compounds of resveratrol, e-viniferin, and δ-viniferin were mainly produced in a 3 L bioreactor culture following elicitation of cells with MJ and STE. These results provide new strategies for conditional, high-level production of resveratrol and viniferins in cell cultures by utilizing the solubilizing properties of STE or CD-M.

Molecular Evaluation of Vitality and Survival Rate of Dormant Kyoho Grape Seedlings: A Step toward Molecular Farming

Vitality and survival rate of grape seedlings are crucial factors affecting quality of vineyards. There is no comprehensive study describing accurate evaluation of dormant grapevine seedlings’ vitality and survival rate. The purpose of this study was to explore the possibility of using molecular information to evaluate viability and survival rate of dormant seedlings before transplanting. After bare roots treatment, 1–5 day expression levels of six HKGs in four buds of tetraploid Kyoho grape (Vitis labruscana: V. labrusca × V. vinifera) seedlings were detected by (Sq.) RT-PCR and qRT-PCR for calibration of the molecular method. The results revealed that HKGs expression indicates vitality and survival of plant, higher expression was strongly linked to higher vitality and survival rate, lower expression was associated with lower vitality, and lowest expression was significantly associated with lowest vitality and survival rate. Moreover, DNA and RNA quality can superficially determine seedling qualities. Finally, the survival rate of the seedlings produced in Juxian-Shandong, Laixi-Shandong, Huailai-Hebei, Suizhong-Liaoning, Changli-Hebei, Guanxian-Shandong, and Zhangjiagang-Jiangsu was 100.00%, 100.00%, 100.00%, 100.00%, 100.00%, 87.77%, and 93.33%, respectively. In conclusion, molecular technique is potential approach for promoting gene information to estimate vitality and survival rate of dormant grape seedlings and might contribute to viticulturists’ efforts.