Citrus asymmetric somatic hybrids produced via fusion of gamma‑irradiated and iodoacetamide‑treated protoplasts

Authors

  • Claudine Maria de Bona IAPAR
  • Jean Howe Gould Texas A&M University - TAMU
  • J. Creighton Miller Jr Texas A&M University - TAMU
  • David Stelly Texas A&M University - TAMU
  • Eliezer Silva Louzada Texas A&M UNiversity Kingsville - TAMUK

DOI:

https://doi.org/10.1590/S1678-3921.pab2009.v44.1832

Keywords:

<i>Citrus</i> sp., gamma irradiation, grapefruit, protoplast fusion, somatic hybridization, sweet orange

Abstract

The objective of this study was to produce citrus somatic asymmetric hybrids by fusing gamma-irradiated protoplasts with iodoacetamide-treated protoplasts. Protoplasts were isolated from embryogenic suspension cells of grapefruit (Citrus paradisi Macfad.) cultivars Ruby Red and Flame, sweet oranges (C. sinensis Osbeck) 'Itaboraí', 'Natal', Valencia', and 'Succari', from 'Satsuma' (C. unshiu Marcow.) and 'Changsha' mandarin (C. reticulata Blanco) and 'Murcott' tangor (C. reticulata x C. sinensis). Donor protoplasts were exposed to gamma rays and receptor protoplasts were treated with 3 mmol L-1 iodoacetamide (IOA), and then they were fused for asymmetric hybridization. Asymmetric embryos were germinated, and the resulting shoots were either grafted onto sour orange, rough lemon or 'Swingle' (C. paradisi x Poncirus trifoliata) x 'Sunki' mandarin rootstock seedlings, or rooted after dipping their bases in indol-butyric acid (IBA) solution. The products were later acclimatized to greenhouse conditions. Ploidy was analyzed by flow cytometry, and hybridity was confirmed by amplified fragment length polymorphism (AFLP) analysis of plantlet DNA samples. The best treatment was the donor-recipient fusion combination of 80 Gy-irradiated 'Ruby Red' protoplasts with 20 min IOA-treated 'Succari' protoplasts. Tetraploid and aneuploid plants were produced. Rooting recalcitrance was solved by dipping shoots' stems in 3,000 mg L-1 IBA solution for 10 min.

Author Biographies

Claudine Maria de Bona, IAPAR

Dr. Claudine Bona, a fruit researcher (pomology, horticulture) of the Agronomic Institute of Parana (IAPAR), Brazil, received her B.S. and M.S. in Agronomy and Plant Sciences from the Federal University of Parana in 1998 and 2002, respectively. She received her Ph.D. in Horticulture from Texas A&M University in 2007 and Post-Doctorate from the Federal University of Parana in 2008. Her research focuses on banana and citrus crop improvement.

Jean Howe Gould, Texas A&M University - TAMU

Adjunct Assistant and Adjunct Professor of the Department of Horticultural Sciences, Jean Gould received her B.A. in Liberal Arts & Sciences from the University of Illinois, Champaign-Urbana and her Ph.D. in Plant Physiology from the University of California, Riverside. Dr. Gould's area of research is genetic engineering and biotechnology.

J. Creighton Miller Jr, Texas A&M University - TAMU

Professor, Department of Horticultural Sciences, Dr. Creighton Miller received his B.S. and M.S. in General Studies and Horticulture from Louisiana State University in 1965 and 1967, respectively. He received his Ph.D. in Horticulture (Breeding and Genetics) from Michigan State University in 1972. Dr. Miller is a Member of the Graduate Faculties of Horticulture, Genetics, and Molecular and Environmental Plant Sciences at Texas A&M University.

David Stelly, Texas A&M University - TAMU

Professor of Plant Breeding, Genetics, Molecular and Environmental Plant Sciences, Dr. Stelly received his B.S. in Genetics from the University of Wisconsin-Madison in 1975, his M.S. in Plant Breeding and Cytogenetics from Iowa State University in 1979 and his Ph.D. in Plant Breeding and Plant Genetics from the University of Wisconsin-Madison in 1983. His research addresses basic and practical needs related to plant reproductive genetics or biology, evolution, chromosome biology and cytogenetic manipulation, genomics, genetic barriers to introgression, or other processes that impact our ability to genetically analyze, manipulate and improve plants.

Eliezer Silva Louzada, Texas A&M UNiversity Kingsville - TAMUK

Assistant Professor of the Citrus Center, at Texas A&M University-Kingsville, in Weslaco, Dr. Louzada is an Adjunct Member of the Graduate Faculty of TAMU. He received his B.S. degree in Agronomy and his M.S. degree in Soil Science from the Federal Rural University of Rio de Janeiro, Brazil in 1975 and 1978. He received his Ph.D. in Genetics in a joint program between the Federal University of Rio de Janeiro and The University of Florida in 1993. Dr. Louzada's research is divided in two areas, breeding, and horticulture. The main focus of his breeding research is to develop a new methodology to allow the transference of a single chromosome between citrus species, between citrus wild relatives and citrus, and between citrus and a short cycle plant.

Downloads

Published

2010-11-18

How to Cite

Bona, C. M. de, Gould, J. H., Jr, J. C. M., Stelly, D., & Louzada, E. S. (2010). Citrus asymmetric somatic hybrids produced via fusion of gamma‑irradiated and iodoacetamide‑treated protoplasts. Pesquisa Agropecuaria Brasileira, 44(5), 454–462. https://doi.org/10.1590/S1678-3921.pab2009.v44.1832

Issue

v.44, n.5, maio 2009

Section

POMOLOGY