Identification of SSR markers for hybridity and seed genetic purity testing in sunflower (Helianthus annuus L.)

Abstract

The genuineness of a hybrid is one of the most important characteristics of good quality seed. In order to identify pure hybrid and pollen shedders/ offtypes an investigation was performed to identify an ideal SSR marker. 58 primer pairs were screened to identify the specific marker associated with each hybrid and parental lines. Hybrid KBSH-44 could be clearly identified by using ORS 309 and ORS 170, based on the banding pattern resolved on polyacrylamide gel (6%). The complementary banding pattern of both parents made a way to identify the hybrid. ORS 309 amplified allele size at 250 bp was specific to female parent (CMS-17A) and 230 bp was specific to male parent (RHA 95-C-1). These two bands of allele size 230 and 250 bp were found only in hybrid KBSH-44. Another SSR primer ORS 170 was able to distinguish the hybrid KBSH-44 by amplifying allele of size 230 bp a female specific (CMS-17A) allele and 200 bp amplicon a male specific allele (RHA 95-C-1). SSR primer ORS 811 found specific to identify KBSH-53 and it amplified allele of size 270 bp in its female parent (CMS-53A) and allele size of 230 bp in its pollen parent (RHA 95-C-1). The hybrid has both the alleles from its parents at 270 and 230 bp.

Optimization of sampling size for DNA-based PCR assay for hybrid purity test in the brinjal, Solanum melongena

“…In addition to capital invested on hybrid seed production and additional expenditure incurred on storage of hybrid seed, GOT ultimately increases the hybrid seed cost (Nandakumar et al, 2004). Similar to present study, use of SSR markers for genetic purity testing has been demonstrated in rice (Nandakumar et al, 2004); in maize (Wang et al, 2002) and in sunfl ower (Pallavi et al, 2011) and overall data remained comparable with fi eld grow out test. Similar to SSR marker applied in present study with minimum sample size; number of workers with other plants have applied different sample size of seeds for RAPD primers for example: 400 seeds in chicory (Bellamy et al, 1998), 120 in canola (Marshall et al, 1994), 40 in tomato (Rom et al, 1995), 30 in Chinese cabbage (Meng et al, 1998) and 10-20 in pepper (Ballester and Vicente, 1998) to assess the genetic purity of the hybrid seeds.…”

A new SNP genotyping technology Target SNP-seq and its application in genetic analysis of cucumber varieties

“…The 261 varieties represented all types and main cucumber varieties planted in China, including 111 commercial hybrid varieties from the Chinese seed market, 67 varieties from breeders' collections in BVRC, 64 varieties from the Chinese government department, and 19 local landraces from Xishuangbanna of Yunnan Province in China (Supplementary Table S2). Previous studies showed that the genetic purity of hybrid seeds is prone to contamination due to the occurrence of out-crossing with foreign or self-pollinated and physical admixtures 36 . In order to increase the genotype accuracy of each cucumber variety, newly expanded young leaves grown from 30 seeds were used for DNA extraction.…”

Grow out test and determination of physical characteristics of seed of rice landraces

“…GOT is time consuming (takes one full growing season for completion), space demanding and often does not allow the unequivocal identification of genotypes. The molecular markers are of great importance for rapid assessment seed purity (Yashitola et al, 2002, Antonova et al, 2006and Pallavi et al, 2011. Rajendrakumar et al (2007) used molecular markers for detection of contaminant in CMS seed stocks of rice and confirm result based on observation recorded in GOT field.…”