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Successful crosses between fungal-resistant wild species of Arachis (section Arachis) and Arachis hypogaea.

Fávero AP, Dos Santos RF, Simpson CE, Valls JF, Vello NA - Genet. Mol. Biol. (2015)

Bottom Line: These sterile hybrids were polyploidized and five combinations produced tetraploid flowers.Next, 16 combinations were crossed between A. hypogaea and the synthetic amphidiploids, resulting in 11 different hybrid combinations.Our results confirm that it is possible to introgress resistance genes from wild species into the peanut using artificial hybridization, and that more species than previously reported can be used, thus enhancing the genetic variability in peanut genetic improvement programs.

View Article: PubMed Central - PubMed

Affiliation: Embrapa Pecuária Sudeste, São Carlos, SP, Brazil.

ABSTRACT
Peanut (Arachis hypogaea) is the fifth most produced oil crop worldwide. Besides lack of water, fungal diseases are the most limiting factors for the crop. Several species of Arachis are resistant to certain pests and diseases. This study aimed to successfully cross the A-genome with B-K-A genome wild species previously selected for fungal disease resistance, but that are still untested. We also aimed to polyplodize the amphihaploid chromosomes; cross the synthetic amphidiploids and A. hypogaea to introgress disease resistance genes into the cultivated peanut; and analyze pollen viability and morphological descriptors for all progenies and their parents. We selected 12 A-genome accessions as male parents and three B-genome species, one K-genome species, and one A-genome species as female parents. Of the 26 distinct cross combinations, 13 different interspecific AB-genome and three AA-genome hybrids were obtained. These sterile hybrids were polyploidized and five combinations produced tetraploid flowers. Next, 16 combinations were crossed between A. hypogaea and the synthetic amphidiploids, resulting in 11 different hybrid combinations. Our results confirm that it is possible to introgress resistance genes from wild species into the peanut using artificial hybridization, and that more species than previously reported can be used, thus enhancing the genetic variability in peanut genetic improvement programs.

No MeSH data available.


Related in: MedlinePlus

Combinations between Arachis hoehnei and A. simpsonii and A. ipaënsis and A. villosa and their progeny. Agarose gel, using LEC primer.
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f01: Combinations between Arachis hoehnei and A. simpsonii and A. ipaënsis and A. villosa and their progeny. Agarose gel, using LEC primer.

Mentions: Leaves from each progeny plant were individually collected and DNA was extracted according to the adapted protocol by Murray and Thompson (1980). The DNA was quantified using agarose gels (1.2%) at 80 V for 1 h and diluted to 2.5 ngmL−1 concentration. The PCR reaction for DNA amplification contained the following reagent cocktail: PCR buffer (10 mM Tris-HCl, pH 8.3, and 50 mM KCl), 1.5 mM MgCl2, 2.5 mM dNTPs, 5 pmol of each primer pair (A1-041, 5′-CGCCACAAGATTAACAAGCACC-3′ and (F) 5′-GCTGGGATCATTGTAGGGAAGG-3′ (R); A1-558, 5′-TGTGACACCATCAATCAAAGGG-3′ (F) and 5′-CAAAACCCAAATCATCACCACC-3′ (R); and LEC-1, (AT) 5′-CAAGCATCAACAACAACGA-3′ (F) and 5′-GTCCGACCACATACAAGAGTT-3′ (R)), 5 U Taq DNA Polymerase, and 10 mg mL−1 BSA (bovine serum albumine), which was mixed separately from DNA; then 2.5 ng mL−1 DNA was added; and Milli-Q sterile water was added to complete the reaction volume to 13 μL. Mineral oil (50 μL) was added to prevent the cocktail from evaporating. The amplification reaction conditions were as follows: denaturation for 5 min at 94 °C; 29 cycles of 1 min at 94 °C, 1 min at 56 °C (60 °C for primer pairs A1-041 and A1-558), and 1 min at 72 °C; and a final extension of 7 min at 72 °C. The amplified products were separated by electrophoresis in 4% agarose gel (9 pv−1), using TBE buffer (0.09 of Tris; 0.09 M boric acid, and 2 mM EDTA), pH 8.0, at constant 90 V cm−1. The gels were stained with 10 mL ethidium bromide (10 mg mL−1) diluted in 100 mL TBE and visualized under ultraviolet light (GELDOC 2000, BIORAD). Figure 1 presents PCR amplifications using the LEC primer pair for two hybrid combinations between A-genome wild species (A. hoehnei - KG 30006 and A. simpsonii V 13710) and A- and B-genome wild species (A. ipaënsis KGBPScS 30076 and A. villosa V 12812), and their progeny.


Successful crosses between fungal-resistant wild species of Arachis (section Arachis) and Arachis hypogaea.

Fávero AP, Dos Santos RF, Simpson CE, Valls JF, Vello NA - Genet. Mol. Biol. (2015)

Combinations between Arachis hoehnei and A. simpsonii and A. ipaënsis and A. villosa and their progeny. Agarose gel, using LEC primer.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4612608&req=5

f01: Combinations between Arachis hoehnei and A. simpsonii and A. ipaënsis and A. villosa and their progeny. Agarose gel, using LEC primer.
Mentions: Leaves from each progeny plant were individually collected and DNA was extracted according to the adapted protocol by Murray and Thompson (1980). The DNA was quantified using agarose gels (1.2%) at 80 V for 1 h and diluted to 2.5 ngmL−1 concentration. The PCR reaction for DNA amplification contained the following reagent cocktail: PCR buffer (10 mM Tris-HCl, pH 8.3, and 50 mM KCl), 1.5 mM MgCl2, 2.5 mM dNTPs, 5 pmol of each primer pair (A1-041, 5′-CGCCACAAGATTAACAAGCACC-3′ and (F) 5′-GCTGGGATCATTGTAGGGAAGG-3′ (R); A1-558, 5′-TGTGACACCATCAATCAAAGGG-3′ (F) and 5′-CAAAACCCAAATCATCACCACC-3′ (R); and LEC-1, (AT) 5′-CAAGCATCAACAACAACGA-3′ (F) and 5′-GTCCGACCACATACAAGAGTT-3′ (R)), 5 U Taq DNA Polymerase, and 10 mg mL−1 BSA (bovine serum albumine), which was mixed separately from DNA; then 2.5 ng mL−1 DNA was added; and Milli-Q sterile water was added to complete the reaction volume to 13 μL. Mineral oil (50 μL) was added to prevent the cocktail from evaporating. The amplification reaction conditions were as follows: denaturation for 5 min at 94 °C; 29 cycles of 1 min at 94 °C, 1 min at 56 °C (60 °C for primer pairs A1-041 and A1-558), and 1 min at 72 °C; and a final extension of 7 min at 72 °C. The amplified products were separated by electrophoresis in 4% agarose gel (9 pv−1), using TBE buffer (0.09 of Tris; 0.09 M boric acid, and 2 mM EDTA), pH 8.0, at constant 90 V cm−1. The gels were stained with 10 mL ethidium bromide (10 mg mL−1) diluted in 100 mL TBE and visualized under ultraviolet light (GELDOC 2000, BIORAD). Figure 1 presents PCR amplifications using the LEC primer pair for two hybrid combinations between A-genome wild species (A. hoehnei - KG 30006 and A. simpsonii V 13710) and A- and B-genome wild species (A. ipaënsis KGBPScS 30076 and A. villosa V 12812), and their progeny.

Bottom Line: These sterile hybrids were polyploidized and five combinations produced tetraploid flowers.Next, 16 combinations were crossed between A. hypogaea and the synthetic amphidiploids, resulting in 11 different hybrid combinations.Our results confirm that it is possible to introgress resistance genes from wild species into the peanut using artificial hybridization, and that more species than previously reported can be used, thus enhancing the genetic variability in peanut genetic improvement programs.

View Article: PubMed Central - PubMed

Affiliation: Embrapa Pecuária Sudeste, São Carlos, SP, Brazil.

ABSTRACT
Peanut (Arachis hypogaea) is the fifth most produced oil crop worldwide. Besides lack of water, fungal diseases are the most limiting factors for the crop. Several species of Arachis are resistant to certain pests and diseases. This study aimed to successfully cross the A-genome with B-K-A genome wild species previously selected for fungal disease resistance, but that are still untested. We also aimed to polyplodize the amphihaploid chromosomes; cross the synthetic amphidiploids and A. hypogaea to introgress disease resistance genes into the cultivated peanut; and analyze pollen viability and morphological descriptors for all progenies and their parents. We selected 12 A-genome accessions as male parents and three B-genome species, one K-genome species, and one A-genome species as female parents. Of the 26 distinct cross combinations, 13 different interspecific AB-genome and three AA-genome hybrids were obtained. These sterile hybrids were polyploidized and five combinations produced tetraploid flowers. Next, 16 combinations were crossed between A. hypogaea and the synthetic amphidiploids, resulting in 11 different hybrid combinations. Our results confirm that it is possible to introgress resistance genes from wild species into the peanut using artificial hybridization, and that more species than previously reported can be used, thus enhancing the genetic variability in peanut genetic improvement programs.

No MeSH data available.


Related in: MedlinePlus