文献类型：期刊文献

英文题名：Shoot tip vitrification cryopreservation and elimination of soybean mosaic virus (SMV) and cauliflower mosaic virus (CMV) from infected stocks of Pinellia Ternata (Thunb.) Breit

作者：Zhang, Yanhong[1];Dong, Wanqi[1];Wang, Jinxiu[1];Gao, Sufang[1];Tang, Shunli[1];Guo, Qingyi[1];He, Chunyu[1]

第一作者：张延红

通信作者：He, CY[1]

机构：[1]Gansu Univ Chinese Med, Coll Pharm, Lanzhou 730000, Peoples R China

第一机构：甘肃中医药大学药学院（西北中藏药协同创新中心办公室）

通信机构：[1]corresponding author), Gansu Univ Chinese Med, Coll Pharm, Lanzhou 730000, Peoples R China.|[1073501e14fb35863569f]甘肃中医药大学药学院（西北中藏药协同创新中心办公室）;[10735]甘肃中医药大学;

年份：2025

卷号：16

期号：1

外文期刊名：3 BIOTECH

收录：;WOS:【SCI-EXPANDED(收录号:WOS:001649405600002)】；

基金：National Natural Science Foundation of China, 81960683. Gansu Provincial University Industry Support Project, 2020C-09.

语种：英文

外文关键词：Pinellia ternata (Thunb.) Breit; Dormant tuber; Cryopreservation; Virus elimination; Genetic stability

摘要：In the present study, a vitrification method for shoot tip cryopreservation and virus eradication in P. ternata is described as follows: After 7 days of cold acclimation of second-generation in vitro tubers in dormancy, shoot tips (0.8-1 mm in length) were excised from in vitro dormant tubers, transferred to cryovials, and treated with Loading Solution (LS) for 20 min. Subsequently, the shoot tips were incubated in Plant Vitrification Solution (PVS2) for 40 min, followed by direct immersion in liquid nitrogen (LN) along with the vitrification solution. After rapid thawing at 38 degrees C for 2 min, the shoot tips were immersed in Unloading Solution for 20 min, then transferred to 1/2 MS medium (half-strength salts) supplemented with Kinetin (KT) 0.5 mgL- 1, alpha-Naphthylacetic acid (NAA) 0.2 mgL- 1, 3% (w/v) sucrose, and 8 g L- 1 agar (pH 5.8) for recovery. Morphologically, plantlets regenerated from cryopreserved shoot tips were identical to non-cryopreserved controls. Inter-Simple Sequence Repeat (ISSR) analysis revealed a mere 0.063% band variation among 44 cryopreserved plantlets. Attempts were also made to eliminate SMV and CMV by shoot tip culture for twice, cryotherapy, and thermotherapy combined with shoot tip culture. All methods received high survival rates and regeneration rates (over 70.0%). Frequency of virus-free plantlets produced by cryotherapy was 85.7% for SMV and 57.1% for CMV, which were higher than by shoot tip culture for twice (80% for SMV and 40% for CMV) and three treatments of thermotherapy followed by shoot tip culture, including 35 degrees C for 4 weeks (88% for SMV and 48% for CMV), 38 degrees C for 2 weeks (85.7% for SMV and 28.0% for CMV) and 35 degrees C for 2 weeks (85.7% for SMV and 14.3% for CMV). This technology can simultaneously support production of virus-free plants and long-term conservation of P. ternata germplasm, which is critical for agricultural production and breeding. Moreover, cryopreservation of shoot tips dissected from dormant buds proves to be a viable strategy for addressing low regeneration rates post-cryogenic treatment.