Molecular and morphological characterization of somatic hybrids between Solanum tuberosum L. and S. etuberosum Lindl.

Abstract

Interspecific potato somatic hybrids between Solanum tuberosum L. (di)haploid C-13 and 1 endosperm balance number non-tuberous wild species S. etuberosum Lindl. were produced by protoplasts electrofusion. The objective was to transfer virus resistance from this wild species into the cultivated potatoes. Post-fusion products were cultured in VKM medium followed by regeneration of calli in MS 13 K medium at 20°C under a 16-h photoperiod, and regenerants were multiplied on MS medium. Twenty-one somatic hybrids were confirmed by RAPD, SSR and cytoplasm (chloroplast/mitochondria) type analysis possessing species-specific diagnostic bands of corresponding parents. Tetraploid nature of these somatic hybrids was determined through flow cytometry analysis. Somatic hybrids showed intermediate phenotypes (plant, leaves and floral morphology) to their parents in glasshouse grown plants. All the somatic hybrids were malefertile. ELISA assay of somatic hybrids after artificial inoculation of Potato virus Y (PVY) infection reveals high PVY resistance.

Analysis of genetic stability of in vitro propagated potato microtubers using DNA markers

“…RAPD primers used in the analysis are summarized in Supplementary Table 1. The polymerase chain reaction (PCR) amplification and analysis was performed as described by Sarkar et al (2011) and Tiwari et al (2010). Inter simple sequence repeat (ISSR) analysis was performed using genomic DNA by ISSR primers summarized in Supplementary Table 1.…”

Protoplast isolation and plant regeneration of different doubled haploid lines of cauliflower (Brassica oleracea var. botrytis)

“…Moreover, plant regeneration from hypocotyls protoplasts of different DH lines of cauliflower was reproducibly achieved and the frequency of shoot regeneration is higher than previously reported for cauliflower, which gave a maximum regeneration rate of 72% (Veera et al 2009). Plant regeneration from protoplasts is influenced by many factors, including the methods of protoplast isolation, inoculation density, culture system, nurse cells, and medium compositions (Meyer et al 2009;Eeckhaut et al 2009;Castelblanque et al 2010;Prange et al 2010;Tiwari et al 2010). The use of feeder layer-culture systems with protoplasts of other species as nurse cells provided a simple and reliable technique for enhancing cell division and plating efficiency of isolated protoplasts (Walters and Earle 1990;Karamian and Ebrahimzadeh 2001).…”

Nuclear DNA content and chromosome number in somatic hybrid allopolyploids of Solanum

“…FCM is also very helpful in the detection of variation in ploidy status among genotypes of the same species, e.g. Brassica napus (Takahira et al 2011), as well as in interspecific hybrids (Tiwari et al 2010). The variability in nuclear genome size of the ngr ?…”