Abstract
CMV is one of the major destructive viruses worldwide and commercial CMV resistant cucumber is very rare. Therefore we aimed to establish the ovule derived resistant lines used for the breeding program. Haploid plants of sixty-eight cucumber lines were successfully obtained by culturing the un pollinated ovaries harvested one day before an thesis on a modified MS medium supplemented with BAP and IAA at the ratio of 2:1 which was optimal to induce embryogenesis in most of the tested lines. For whole plant regeneration, another modified MS medium was used supplemented with a combination of 6-Benzylaminopurine (BAP) and Indole-3-Acetic Acid (IAA) (2:1) or BAP and IAA/6-(gamma, gamma-dimethylallylamino) purine (2ip) and IAA (5:1) and 5 ppm AgNO3. Ploidy levels of the regenerants were determined by cytological analysis. Thirteen out of 42 clones derived from 14 accessions showed a chromosome number of n = 7 and chloroplast number of 6/pair of guard cell, 24 lines were auto-dihaploid with n = 14 and a chloroplast number of 11-12/pair of guard cell. Twenty-eight Double Haploid (DH) lines were mechanically inoculated with CMV and the level of resistance was evaluated by using DAS-ELISA. Ten highly Resistant lines (R) included 70S2, 91e, 91.1, 93S4-1, 93S42-, 95S1-2, 95S2 DHS1, 117S2-1-3, 136.1 and 194S1 did not show any virus symptom and gave negative ELISA results. Twelve moderately resistant clones were identified including two clones from line 11, three clones from line 93, four clones from line 91 and one clone each from line 117S2 and 123, whereas clone 11.4 was moderately susceptible. Five DH clones; 117S2-1-1, 117S2-2, 117S2-4, 117S2-7 and 117S2-8were classified as highly susceptible.
Highlights
Cucumber (Cucumis sativus) is one of the most baths) (OAE, 2014)
Factors affecting the success of the cucumber ovule culture were identified as the genotypes of the donor plants and the hormonal balance of auxin and cytokinin
A modified Murashigeand Skoog (MS) medium supplemented with BAP and Indole-3-Acetic Acid (IAA) at the ratio of 2:1 was optimal for the ovule culture of most cucumber lines used in the current research and embryo induction was successfully obtained by transfer of the cultured ovules to MS plus BAP and IAA or 2ip and IAA at the ratio of 5:1
Summary
Cucumber (Cucumis sativus) is one of the most baths) (OAE, 2014). The OAE data, indicates that the economically important vegetable crops in Thailand where it is grown intensively all year around. In 2012, a total of 87.82 tons was exported to (CMV) is one of the important viruses in temperate, tropic and sub tropic regions of the world which could cause crop. CMV is one of the most widely spread virus in the world infecting over 1,000 plant species belonging to more than 85 families (Roossinck, 2002). This virus is transmitted by several species of aphid in a nonpersistent manner which is an easy way to transmit the virus from the infected plant to healthy ones. Homozygous double haploid lines from new cucumber accessions could be used to accelerate breeding for new resistant commercial varieties. The purpose of this study are to establish Double Haploid (DH) plant lines by ovule culture and to screen DH lines
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