Abstract
BackgroundArachis hypogaea L. (2n = 4× = 40, AABB) is one of the most important oil and economic crop plants in the word. This species has the largest genome size of about 2,813 Mb among the oil crop species. Zhonghua 8 is a peanut cultivar planted widely in central China and has several superior traits including high oil content, high yield and disease resistance. A high-quality BAC library of Zhonghua 8 was constructed for future researches on the genomics of Chinese peanut cultivars.ResultsA Hin d III-digested genomic BAC (bacterial artificial chromosome) library was constructed with the genomic DNA from leaves of Zhonghua 8. This BAC library consists of 160,512 clones and the average insert is estimated about 102 kb ranging from 30 to 150 kb. The library represents about 5.55× haploid genome equivalents, and provides a 99.71% probability of finding specific genes. The empty-vector rate is under 5 percent detected from 200 randomly selected clones. Probing of 384 clones with the psbA gene of barley chloroplast and the atp6 gene of rice mitochondrion indicated that the contamination with organellar DNA is insignificant. Successive subculture of three clones showed that the inserts are stable in one hundred generations.ConclusionsThis study presented the construction of a high-quality BAC library for the genome of Chinese cultivated peanut. Many essential experiences were summarized in the present study. This BAC library can serve as a substantial platform for development of molecular marker, isolation of genes and further genome research.Electronic supplementary materialThe online version of this article (doi:10.1186/1999-3110-55-8) contains supplementary material, which is available to authorized users.
Highlights
Arachis hypogaea L. (2n = 4× = 40, AABB) is one of the most important oil and economic crop plants in the word
The following steps were only followed for high-molecular weight (HMW) DNA extraction: the plugs were incubated at 50°C for 24 h in lysis buffer [0.005 M citric acid, 0.14 M NaCl, 0.05 M Na2EDTA, 2% (w/v) PVP-40, 1% (w/v) sodium dodecyl sulfate (SDS), 1% sodium lauryl sarcosine titrated to pH 6.5 with NaOH, and autoclaved] and the same antioxidants at similar proportions as in the extraction buffer, and 0.2 mg/ ml Proteinase K was added
Production of Bacterial artificial chromosome (BAC) library Eleven successful ligation reactions were performed for construction of the BAC library
Summary
(2n = 4× = 40, AABB) is one of the most important oil and economic crop plants in the word. This species has the largest genome size of about 2,813 Mb among the oil crop species. Cultivated peanut (Arachis hypogaea L.), a species of the genus Arachis, is one of the most important oilseed crops in tropical and subtropical regions of the world, mainly planted in India, China, Nigeria, Senegal, Sudan, and other countries. Section Arachis is the largest section and harbors the only cultivated peanut species A. hypogaea that is very probably derived from a unique cross between the wild diploid species A. duranensis (A-genome) and A. There is a significant need to pursue genomic strategies in cultivated peanut, with the specific goal of clarifying the genomic construction of different peanut cultivars
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