Development of ent-kaurene Oxidase-Based Conserved Intron Spanning Primers for Species Identification in the Genus Poa (Poaceae; Bluegrass)

Jonathan Lamantia,David Huff,Ambika Chandra

doi:10.3390/agronomy8050058

Abstract

Interspecific hybridization has been attempted to combine the heat and drought of Poa arachnifera Torr. with the turf quality characteristics of several Poa species. Confirmation of an F1 hybrid through morphological analysis of vegetative and flowering characteristics is often time consuming and ambiguous. Ent-kaurene oxidase (KO) has been sequenced in rice, barley, and wheat. In rice, each of the five copies of KO gene has unique lengths for the first intron. Conserved intron spanning primers (CISP) can be used as a DNA marker to exploit variations of intron lengths that flank conserved gene sequences. In the present study, we developed CISP to sequence partial genomic fragments of the KO gene from seven Poa species. Through sequence analysis, species-specific primers were also developed to produce co-dominant markers that can be used to identify interspecific hybrids between Texas bluegrass and six other Poa species used in the present study.

Highlights

Texas bluegrass (Poa arachnifera Torr.) is a dioecious, cool-season grass with exceptional heat and drought tolerance; it lacks higher levels of tiller density and a finer leaf texture that are desirable for residential and athletic field applications [1]
The primer pair kaurene oxidase (KO)-F/KO-R allowed for the amplification of the KO homolog from all seven Poa species used in the present study
P. compressa, P. supina, and P. trivialis each produced a single amplicon ranging in lengths from 1405 bp to 1383 bp while P. annua, P. arachnifera, P. infirma, and P. pratensis each produced multiple amplicons ranging in lengths from 1538 bp to 1327 bp (Table 1)

Summary

Introduction

Texas bluegrass (Poa arachnifera Torr.) is a dioecious, cool-season grass with exceptional heat and drought tolerance; it lacks higher levels of tiller density and a finer leaf texture that are desirable for residential and athletic field applications [1]. The hybridization of Texas bluegrass and Kentucky bluegrass can bring together the traits of heat and drought tolerance with increased turf quality and apomixis [3,4,5]. Presl) have been hybridized with Texas bluegrass as an alternative means to increase turf quality, abiotic stress tolerance, and sexual reproduction and to allow for breeding of desirable traits in advanced generations [1,6,7]. Confirmation of hybrids can be done through analysis of morphological characteristics; molecular markers would greatly increase the efficiency of the breeding process

Methods

Results

Discussion

Conclusion