BGN 19: Comparative inheritance of hordeins in doubled haploids of barley obtained by the Hordeum bulbosum and anther culture techniques

Comparative inheritance of hordeins in doubled haploids of barley obtained by the Hordeum bulbosum and anther culture techniques

P. Devaux
Florimond Desprez Seeds Co.
POB 41, F-59242 Cappelle en Pevele
France

F. Huebner
Northern Regional Research Center, USDA-ARS
1815 N. University St.
Peoria, IL 61604 USA

T. Burnouf
C.R.T.S., F-59012
Lille Cedex
France and

J. Bietz
Northern Regional Research Center, USDA-ARS
1815 N. University St.
Peoria, IL 61604 USA


Doubled haploids (DHs) of barley are frequently produced either by the Hordeum bulbosum or by anther culture techniques (Devaux, 1987). Both methods are amenable to barley breeding programs, but their respective advantages for plant production efficiency or genetic transmission of characters are little known. The present study compares the genetic inheritance of hordeins, the major storage proteins of barley, in microspore- (MDH) and H. bulbosum- (HbDH) doubled haploid lines derived from F1 hybrids from parents having different B and C hordeins. Hordeins were analyzed by reversed-phase high-performance liquid chromatography (RP-HPLC), an excellent method of cereal protein fractionation (Bietz, 1983). The RP-HPLC absorption patterns for the parents and some progeny are shown in Figures I and 2.

No recombination was found between Hor I and Hor 2 loci, coding C and B hordeins, respectively, in the MDH. By contrast, 7 out of 49 (14.3%) HbDH analyzed were recombinants (Table 1). This latter rate of recombination between these two loci is similar to that reported by Jensen (1987), while Shewry et al. (1978) found higher recombination percentages in male than in female gametes.

Table 1. Numbers of doubled haploids (DH) showing qualitatively identical C and B hordein patterns (NR=Non-Recombinant) and DHs which have inherited C hordeins from one parent and B hordeins from the other (R=Recombinant).

Fig. 1. RP-HPLC separations of C and B hordeins (early- and late- eluting, respectively) from the parents DH8293 (1) and Kaskade (2); from microspore-derived doubled haploid lines from the Fi hybrid (C163) of these parents which are qualitatively identical to one of the parents [(3) 1-6a; (4) 2-4b]; and from a H. bulbosum-derived doubled haploid line from the F1 hybrid which has inherited C hordein from one parent and B hordein from the other [(5) 12-1b].

Fig. 2. RP-HPLC separations of C and B hordeins (early- and late- eluting, respectively) from the parents FD16 (1) and FD21 (2); from microspore-derived doubled haploid lines from the F1 hybrid (C220) of these parents which are qualitatively identical to one or the other parent [(3) 9-6a; (4) 9-2a]; and from H. bulbosum-derived doubled haploid lines from the Fi hybrid which have inherited C hordein from one parent and B hordein from the other [(5) 9-5b; (6) 10-6a].

Studies are in progress to determine the percentage of recombination in classical breeding lines from the same crosses.

These preliminary results suggest that the H. bulbosum technique, as compared to the anther culture technique, could significantly favor obtaining recombinant DHs as far as hordein composition is concerned. This could also show that the recombination percentage is not the same for the male as for the female side. It remains to be seen whether the same phenomenon occurs in other crosses and/or for other characters.

This study also demonstrates that RP-HPLC is a very good tool for genetic studies of barley and other crop species in breeding programs.

References:

Bietz, J.A. 1983. Separation of cereal proteins by reversed-phase high- performance liquid chromatography. J. Chromatogr. 255:219-239.

Devaux, P. 1987. Comparison of anther culture and Hordeum bulbosum method for the production of doubled haploid in winter barley. I. Production of green plants. Plant Breeding 98:215-219.

Jensen, J. 1987. Coordinators' report: Chromosome 5. BGN 17:111-113.

Shewry, P.R., H.M. Pratt, R.A. Finch, and B.J. Miflin. 1978. Genetic analysis of hordein polypeptides from single seeds of barley. Heredity 40:463-466.


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