Romadanova Natalya V, Mishustina Svetlana A, Gritsenko D ilyara A, Omasheva Madina Y, Galiakparov Nurbol N, Reed Barbara M, Kushnarenko Svetlana V
Institute of Plant Biology and Biotechnology, Almaty, Republic of Kazakhstan.
United States Department of Agriculture, Agricultural Research Service, National Clonal Germplasm Repository, Corvallis, OR, USA.
Cryo Letters. 2016 Jan-Feb;37(1):1-9.
There is an urgent need in Kazakhstan for virus-free nursery stock to reinvigorate the industry and preserve historic cultivars. An in vitro collection of apples could be used for virus testing and elimination and to provide virus-free elite stock plants to nurseries.
Malus sieversii Ledeb. M. Roem. and Malus domestica Borkh. accessions were initiated in vitro for virus identification and elimination. Reverse transcription and multiplex PCR were used to test for five viruses. PVS2 vitrification was used as a tool for cryotherapy.
Four viruses, Apple chlorotic leaf spot virus (ACLSV), Apple stem pitting virus (ASPV), Apple stem grooving virus (ASGV) and Apple mosaic virus (ApMV) were detected in 17 accessions. Tomato ringspot virus (ToRSV) was not detected. ACLSV affected 53.8% of the accessions, ASPV 30.8%, ASGV 5.1%, and ApMV was found only in 'Aport Alexander'. Cryotherapy produced virus-free shoot tips for seven of nine cultivars tested. Six cultivars had 60-100% elimination of ACLSV.
An in vitro collection of 59 accessions was established. Virus elimination using cryotherapy produced virus-free shoots for seven of nine cultivars and is a promising technique for developing a virus-free apple collection.
哈萨克斯坦迫切需要无病毒苗木来振兴苹果产业并保存历史悠久的品种。苹果的离体种质库可用于病毒检测和清除，并为苗圃提供无病毒的优良砧木。
对塞威士苹果（Malus sieversii Ledeb. M. Roem.）和苹果（Malus domestica Borkh.）品种进行离体培养以进行病毒鉴定和清除。采用逆转录和多重聚合酶链式反应（PCR）检测五种病毒。利用PVS2玻璃化法进行冷冻疗法。
在17个品种中检测到四种病毒，即苹果褪绿叶斑病毒（ACLSV）、苹果茎痘病毒（ASPV）、苹果茎沟病毒（ASGV）和苹果花叶病毒（ApMV）。未检测到番茄环斑病毒（ToRSV）。ACLSV感染了53.8%的品种，ASPV为30.8%，ASGV为5.1%，ApMV仅在“Aport Alexander”品种中被发现。冷冻疗法为九个测试品种中的七个产生了无病毒茎尖。六个品种对ACLSV的清除率达到60%-100%。
建立了一个包含59个品种的离体种质库。利用冷冻疗法清除病毒为九个品种中的七个产生了无病毒嫩枝，是开发无病毒苹果种质库的一项有前景的技术。
