This paper focuses on the creation of an in vitro collection of grapevine hybrids from the breeding program of the Kazakh Scientific Research Institute of Fruit Growing and Viticulture and investigates the presence of Plasmopara viticola resistance mediated by Rpv3 and Rpv12 loci. We looked at the optimization of in vitro establishment using either shoots taken directly from field-grown plants or from budwood cuttings forced indoors. We further screened for the presence of endophyte contamination in the initiated explants and optimized the multiplication stage. Finally, the presence of the resistance loci against P. viticola was studied. The shoots initiated from the field-sourced explants were the more effective method of providing plant sources for in vitro initiation once all plant accessions met the goal of in vitro establishment. The concentration of phytohormones and the acidity of the culture medium have a great effect on the multiplication rate and the quality of in vitro stock cultures. Out of 17 grapevine accessions, 16 showed the presence of single or combined resistance loci against P. viticola. The grapevine accessions identified as carrying Rpv3 and Rpv12 alleles represent important genetic resources for disease resistance breeding programs. These accessions may further contribute to the creation of new elite cultivars of economic interest.

The loss of plant species, especially endangered and endemic ones, poses a significant threat to global biodiversity. These species cannot easily be replaced when their populations decline or become extinct, which makes their loss particularly devastating. This study focuses on the geobotanical study of nine Rosaceae species (Cotoneaster karatavicus, Crataegus ambigua, Malus niedzwetzkyana, Malus sieversii, Prunus tenella, Prunus ulmifolia, Sibiraea laevigata, Sorbus persica, and Spiraeanthus schrenkianus) and the development of ex situ approaches for the conservation of Rosaceae species listed in the Red Book of Kazakhstan. The geobotanical study revealed an alarming trend of biodiversity loss in five regions of Kazakhstan. This loss is driven by threats from diseases and pests, as well as the aging of plants, small population sizes, weak in situ fruiting, and other factors, such as climate change. We have established an in vitro collection for the short- and medium-term conservation of seeds, embryos and shoots taken either directly from field-grown plants or from budwood cuttings forced indoors. We also use long-term sexual conservation methods, such as the cryopreservation of seed and embryonic axes, alongside conventional seed banking at −20 °C. Ex situ conservation efforts that use multiple propagules and storage methods for the same species are well-suited to a diverse genebank facility. These efforts enable future generations to use this valuable reservoir of genetic diversity for crop improvement and may also serve as a basis for propagating planting material to restore degraded populations.

Tissue culture methods enable virus elimination from vegetatively propagated crop plants but cannot prevent new infections. Here we used a tissue culture transgenic approach for curing field cultivars of Solanum tuberosum through the stimulation of RNA interference (RNAi)-based antiviral defenses. Expression cassettes carrying inverted repeats of potato virus S (PVS, genus Carlavirus) movement or coat protein sequences were used for the transformation of potato cultivars naturally infected with PVS and/or a related carlavirus potato virus M (PVM), without or with potato virus Y (PVY, genus Potyvirus). A high proportion of transformants PCR-positive for transgenes were cured from both carlaviruses and PVY. After 3-year field trials, 22 transgenic lines representing seven cultivars remained free of any virus or became infected only with PVY. Vegetative progenies of the transgenic lines of cultivar Zeren (initially coinfected with PVS, PVM, and PVY), sampled after in vitro propagation or field trials, and other field cultivars accumulated transgene-derived 21, 22, and 24 nt small interfering (si)RNAs almost exclusively from the PVS inverted repeats. Additionally, some field progenies accumulated 21–22 nt siRNAs from the entire PVY genome, confirming PVY infection. Taken together, transgenic RNAi is effective for virus elimination from naturally infected potato cultivars and their sequence-specific immunization against new infections.