Identification of Amplified Fragment Length Polymorphism (AFLP) Markers Tightly Associated with Drought Stress Gene in Male Sterile and Fertile Salvia miltiorrhiza Bunge

Abstract

Consistent grain yield in drought environment has attracted wide attention due to global climate change. However, the important drought-related traits/genes in crops have been rarely reported. Many near-isogenic lines (NILs) of male sterile and fertile Salvia miltiorrhiza have been obtained in our previous work through testcross and backcross in continuous field experiments conducted in 2006–2009. Both segregating sterile and fertile populations were subjected to bulked segregant analysis (BSA) and amplified fragment length polymorphism (AFLP) with 384 and 170 primer combinations, respectively. One out of 14 AFLP markers (E9/M3246) was identified in treated fertile population as tightly linked to the drought stress gene with a recombination frequency of 6.98% and at a distance of 7.02 cM. One of 15 other markers (E2/M5357) was identified in a treated sterile population that is closely associated with the drought stress gene. It had a recombination frequency of 4.65% and at a distance of 4.66 cM. Interestingly, the E9/M3246 fragment was found to be identical to another AFLP fragment E11/M4208 that was tightly linked to the male sterile gene of S. miltiorrhiza with 95% identity and e-value 4 × 10−93. Blastn analysis suggested that the drought stress gene sequence showed higher identity with nucleotides in Arabidopsis chromosome 1–5.

Improvements in the detection power of algorithms for analyzing next-generation sequencing based bulked segregant analysis data via estimating thresholds at the genomic region level

“…Thus the bulk and parent-specific markers should be associated with the trait. Initially, the bulks were genotyped using technologies such as restriction fragment length polymorphism (RFLP) and amplified fragment length polymorphism (AFLP) (Asnaghi et al, 2004;Cai et al, 2003;Hartl et al, 2011;Zhang et al, 2013;Michelmore et al, 1991;Giovannoni et al, 1991). This approach relies on the absence/presence of trait-associated DNA fragments in the gel, and it works well for traits controlled by a single qualitative gene.…”

Physiological and transcriptome analysis reveal molecular mechanism in Salvia miltiorrhiza leaves of near-isogenic male fertile lines and male sterile lines

“…The S. miltiorrhiza cultivation is, however, affected by the factors such as climate and region, which lead to unstable quality of active ingredients. Since the discovery of the male sterile mutants of S. miltiorrhiza , we have created and developed near-isogenic male fertile lines (MF) and near-isogenic male sterile lines (MS) through multiple consecutive testcrosses and backcrosses [20]. It promises possibility to achieve stable and controllable quality of active ingredients.…”

“…Since the discovery of the male sterile mutants of S. miltiorrhiza in 2002, our team has done some research work [7, 20]. Researchers have studied the development and biological properties of pollen.…”

Physiological and transcriptome analysis reveal molecular mechanism in Salvia miltiorrhiza leaves of near-isogenic male fertile lines and male sterile lines

“…It provides us with the possibility to achieve stable and controllable quality of active ingredients. Since the discovery of the male sterile mutant of S. miltiorrhiza, we have created and developed near-isogenic male fertile lines (MF) and near-isogenic male sterile lines (MS) through multiple consecutive testcrosses and backcrosses [16]. The genetic linkage map between male fertile lines and male sterile lines of the S. miltiorrhiza mutant was constructed by amplified fragment length polymorphism (AFLP).…”