Effects of mesoclimate and microclimate variations mediated by high altitude and row orientation on sucrose metabolism and anthocyanin synthesis in grape berries

Climate change and extreme weather pose significant challenges to the traditional viticulture regions.Emerging high-altitude grape-producing regions with diverse orientations have shown great potential in coping with this challenge.Stable,high-quality wine grape production may be achieved by synchronizing the meso-and microclimate.To clarify the role of high altitude and row orientation in meso-and microclimate and the response of berries to it,we evaluated seven years(2012-2018)of climate data,two years of basic grape(Cabernet Sauvignon,Vitis vinifera L.)quality,and one-year microclimate from veraison to harvest.By comparing two locations(Sidon 2047 m,Sinon 2208 m)in Yunnan Province,China,we found that the average temperature has been stable at approximately 15℃ for seven years,with no extreme weather or,noticeable global warming.The light intensity(LI)in the north-south(NS)was more balanced than the east-west(EW)direction,and the east-west to the south(EW-S)canopy side was almost higher than the other sides.High LI was associated with high photosynthetically active radiation(PAR),ultraviolet(UV),and infrared(IR)light and vice versa.The north-south to the east(NS-E)and east-west to the north(EWN)sides were characterized by lower LI and higher UV and IR light,and higher total anthocyanin content.Most anthocyanin synthesis-related genes,for example,VvF3'H and VvF3'5'H,were highly expressed in NS-E from veraison to maturity.Perhaps UV and IR light induced their expression.This study provides new insights on the role of differently orientated rows in controlling grape quality due to varied light quality.The findings are globally significant,particularly in the context of climate change,and offer fresh insights into berry physiological responses and decision-making for the management of existing vineyards.

Whole-genome sequencing reveals rare off-target mutations in CRISPR/Cas9-edited grapevine

The CRISPR(clustered regularly interspaced short palindromic repeats)-associated protein 9(Cas9)system is a powerful tool for targeted genome editing,with applications that include plant biotechnology and functional genomics research.However,the specificity of Cas9 targeting is poorly investigated in many plant species,including fruit trees.To assess the off-target mutation rate in grapevine(Vitis vinifera),we performed whole-genome sequencing(WGS)of seven Cas9-edited grapevine plants in which one of two genes was targeted by CRISPR/Cas9 and three wild-type(WT)plants.In total,we identified between 202,008 and 272,397 single nucleotide polymorphisms(SNPs)and between 26,391 and 55,414 insertions/deletions(indels)in the seven Cas9-edited grapevine plants compared with the three WT plants.Subsequently,3272 potential off-target sites were selected for further analysis.Only one off-target indel mutation was identified from the WGS data and validated by Sanger sequencing.In addition,we found 243 newly generated off-target sites caused by genetic variants between the Thompson Seedless cultivar and the grape reference genome(PN40024)but no true off-target mutations.In conclusion,we observed high specificity of CRISPR/Cas9 for genome editing of grapevine.