Construction and characterization of a bacterial artificial chromosome library for the hexaploid wheat line 92R137.

Biomed Res Int

State Key Laboratory of Crop Stress Biology for Arid Areas and College of Plant Protection, Northwest A&F University, Yangling, Shaanxi 712100, China.

Published: November 2015

For map-based cloning of genes conferring important traits in the hexaploid wheat line 92R137, a bacterial artificial chromosome (BAC) library, including two sublibraries, was constructed using the genomic DNA of 92R137 digested with restriction enzymes HindIII and BamHI. The BAC library was composed of total 765,696 clones, of which 390,144 were from the HindIII digestion and 375,552 from the BamHI digestion. Through pulsed-field gel electrophoresis (PFGE) analysis of 453 clones randomly selected from the HindIII sublibrary and 573 clones from the BamHI sublibrary, the average insert sizes were estimated as 129 and 113 kb, respectively. Thus, the HindIII sublibrary was estimated to have a 3.01-fold coverage and the BamHI sublibrary a 2.53-fold coverage based on the estimated hexaploid wheat genome size of 16,700 Mb. The 765,696 clones were arrayed in 1,994 384-well plates. All clones were also arranged into plate pools and further arranged into 5-dimensional (5D) pools. The probability of identifying a clone corresponding to any wheat DNA sequence (such as gene Yr26 for stripe rust resistance) from the library was estimated to be more than 99.6%. Through polymerase chain reaction screening the 5D pools with Xwe173, a marker tightly linked to Yr26, six BAC clones were successfully obtained. These results demonstrate that the BAC library is a valuable genomic resource for positional cloning of Yr26 and other genes of interest.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4026951PMC
http://dx.doi.org/10.1155/2014/845806DOI Listing

Publication Analysis

Top Keywords

hexaploid wheat
12
bac library
12
bacterial artificial
8
artificial chromosome
8
wheat 92r137
8
765696 clones
8
hindiii sublibrary
8
bamhi sublibrary
8
clones
6
library
5

Similar Publications

This study examines the complex interactions between wheat cultivar selection and fortification with NaFeEDTA and ascorbic acid (AA) on the bioavailability of iron (Fe) and zinc (Zn) in whole wheat flour (WWF) and chapati. Nineteen hexaploid wheat cultivars were rigorously assessed for their intrinsic Fe and Zn profiles, including total content (TC), solubility (S), and bio-accessibility (B), utilizing an in-vitro gastrointestinal model. Significant variations (P < 0.

View Article and Find Full Text PDF

This study focused on identifying amylase-trypsin inhibitors (ATIs) in seven Norwegian-cultivated wheat varieties, including common wheat and ancestral species, and identifying potentially harmful opioid peptides within the digesta of these wheats. LC-MS/MS analysis of tryptic peptides from ATI fractions revealed that the common wheat variety Børsum exhibited the highest diversity of ATIs ( = 24), while they were less represented in tetraploid emmer ( = 11). Hexaploid wheat Bastian showed low diversity and relative abundance of ATIs.

View Article and Find Full Text PDF

Drought, as an abiotic stressor, globally limits cereal productivity, leading to early aging of leaves and lower yields. The expression of the isopentenyl transferase (IPT) gene, which is involved in cytokinin (CK) biosynthesis, can delay drought-induced leaf senescence. In this study, the Agrobacterium Isopentenyl transferase (IPT) gene was introduced into two local hexaploid wheat cultivars, NR-421 and FSD-2008.

View Article and Find Full Text PDF

GBS read coverage analysis identified a Robertsonian chromosome from two Thinopyrum subgenomes in wheat, conferring leaf and stripe rust resistance, drought tolerance, and maintaining yield stability. Agropyron glael (GLAEL), a Thinopyrum intermedium × Th. ponticum hybrid, serves as a valuable genetic resource for wheat improvement.

View Article and Find Full Text PDF

Synthetic intergeneric amphydiploids and genome-substituted wheat forms are an important source for transferring agronomically valuable genes from wild species into the common wheat (Triticum aestivum L.) genome. They can be used both in academic research and for breeding purposes as an original material for developing wheat-alien addition and substitution lines followed by translocation induction with the aid of irradiation or nonhomologous chromosome pairing.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!