Bitencourt, G. A.

Chiari, Lucimara

Leguizámon, G. O. C.

Salgado, L. R.

Valle, Cacilda B.

English

University of Kentucky

University of Kentucky UKnowledge International Grassland Congress Proceedings XXI International Grassland Congress / VIII International Rangeland Congress Use of RAPD Markers to Identify Intraspecific Hybrids of Brachiaria humidicola Cacilda B. Valle EMBRAPA, Brazil Lucimara Chiari EMBRAPA, Brazil G. A. Bitencourt EMBRAPA, Brazil L. R. Salgado EMBRAPA, Brazil G. O. C. Leguizámon EMBRAPA, Brazil Follow this and additional works at: https://uknowledge.uky.edu/igc  Part of the Plant Sciences Commons, and the Soil Science Commons This document is available at https://uknowledge.uky.edu/igc/21/12-2/36 The XXI International Grassland Congress / VIII International Rangeland Congress took place in Hohhot, China from June 29 through July 5, 2008. Proceedings edited by Organizing Committee of 2008 IGC/IRC Conference Published by Guangdong People's Publishing House This Event is brought to you for free and open access by the Plant and Soil Sciences at UKnowledge. It has been accepted for inclusion in International Grassland Congress Proceedings by an authorized administrator of UKnowledge. For more information, please contact UKnowledge@lsv.uky.edu. 　 Multifunctional Grasslands in a Changing World 　 Volume Ⅱ 　 瞯 ]417　 瞯Grasslands/Rangelands Production Systems——— Developing Improved PlantsUse of RAPD markers to identify intraspecific hybrids of Brachiaria humidicolaC .B . V alle , L . Chiari , G .A . Bitencourt , L .R . Salgado , G .O .C . LeguizámonEmbrapa Bee f Cattle , BR 262 km 4 , Cx . Postal 154 , CEP 79002‐970 , Campo Grande , MS , Brasil . E‐mail : cacilda＠cnp gc .embrapa .brKey words : brachiaria grasses , hybrids , molecular markers , koronivia grass , tropical foragesIntroduction Brachiaria humidicola , also known as koronivia grass , is an African species well adapted to acid and poorlydrained soils . It covers millions of hectares of pastures in tropical Latin America and it is used both for cattle and horses .Breeding was done for the first time in this species at Embrapa Beef Cattle , once a sexual tetraploid ecotype was identified in acollection of ５８ accessions of B . humidicola introduced from eastern Africa . Controlled pollination in the greenhouse yieldedhybrids from a cross between the sexual genotype and an apomictic accession . Since no emasculation was done , identification ofhybrids vs . selfs became crucial thus RAPD markers were developed to discriminate hybrids precisely and early in the program .Material and methods Hybridization between the sexual tetraploid accession H３１ of B . humidicola and apomictic cultivar BRSTupi ( H１６) produced ３４８ plants . Seeds were collected on the sexual plant only . DNA was extracted from leaves using theprotocol established by Bonato et al . ( ２００２ ) . Ten randomic primers were selected based on discrimination between the twoprogenitors . The selection criterion was the presence of informative bands , i .e . , those present in the apomictic parent andabsent in the female parent . The identification of hybrids was done directly on the agarose gels analyzed . The presence of atleast three informative bands characterized a hybrid genotype .Results The ten selected primers amplified ３１ informative markers . Primer AB０１ was the one that amplifies most bands ( Table１ ) . Among the plants analyzed , ２８５ presented bands from the apomictic parent ( red arrows ) , which confirms their hybridnature whereas ６３ (１８ .１％ ) did not present any of those bands and were considered results from self‐pollination . These can bediscarded from the breeding program while the others are being phenotyped for mode of reproduction to study inheritance ofapomixis in B . humidicola . Figure １ exemplifies the RAPD profiles generated with primer AB０１ in the parental material ( ApoH１６ and Sex H３１) and in １７ plants from the population .Table 1 Sequence o f nucleotides ( SN ) o f the p rimers used and the number o f in f ormative markers ( IM ) amp li f ied perp rimer .Primer SN IM Primer SN IMP１４  CCAGCCGAAC ５ 儍AC１７ CCTGGAGCTT ２ 寣８１ 儋GGAGCGTACT ３ 儍AD０１  CAAAGGCCGG ３ 寣AB０１ WCCGTCGGTAG ６ 儍AK０４  AGGGTCGGTC ２ 寣BA０２ WTGCTCGGCTC １ 儍AK１９  TCGCAGCGAG ５ 寣AB０２ WGGAAACCCCT １ 儍AL０３  CCCACCCTTG ３ 寣Figure 1 . RA PD p ro f ile o f the parental material and 17 p lants f rom the population under study using the A B01 p rimer .Conclusions RAPD markers adequately selected allow for early identification of hybrids . The approximate proportion of hybridplants to those resulting from selfing was ５ :１ . B . humidicola , and allogamous plant , withstands selfing when subjected toartificial pollination in the greenhouse .ReferenceBonato ,A .L .V . ,Valle ,C .B . ,Jank ,L . ,Resende ,R .M .S . ,Leguizámon ,G .O .C . ２００２ . Extra毕本o de DNA gen蹦mico deBrachiaria e Panicum maximum . Campo Grande , MS : Embrapa Gado de Corte . ( Embrapa Gado de Corte . ComunicadoTécnico ,７９) .

Use of RAPD Markers to Identify Intraspecific Hybrids of \u3ci\u3eBrachiaria humidicola\u3c/i\u3e

https://uknowledge.uky.edu/cgi/viewcontent.cgi?article=3341&amp;context=igc

Use of RAPD Markers to Identify Intraspecific Hybrids of \u3ci\u3eBrachiaria humidicola\u3c/i\u3e

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