The seeds of pollen regenerant And44 after the first (And144) and the second (And244) self-pollinations and the seeds of the donor hybrid H99xWf9 and its parental forms, lines H99 and Wf9, served as a material for electrophoresis. Electrophoretical analysis of storage proteins, zeins, was conducted in flat polyacrylamide gel.

Pollen regenerant And44 was obtained through anther culture of hybrid H99xWf9, colchicinated and planted into the soil. After the self-pollination of this regenerant we had obtained twelve grains of And144, eleven of which were analysed electrophoretically, and one was again planted in the field. After the self-pollination of this F1-plant and the getting of And244 - grains, part of them (fifteen grains) once more were taken for electrophoresis of storage proteins, and another part was anew planted in the field. The plants of lines H99 and Wf9 were also grown in the field for morphobiological investigations.

Electrophoretical spectra of zeins for all the analysed And144 grains (the first generation) and all the analysed And244 grains (the second generation) were the same. This means absence of segregation on given characteristics in the first and the second generations.

The electrophoretical spectra of the regenerant, its donor hybrid H99xWf9 and its parental lines H99 and Wf9 are shown in the figure. The spectrum of the regenerant was not identical to the spectrum of the donor hybrid. Differences in the intensity of the three lines were observed (1-3). One of the lines which was double in donor hybrid spectrum was single in the regenerant (4). The line in the upper part of the hybrid spectrum was absent in the regenerant one (5). The regenerant has some lines which were absent in the hybrid spectrum (6).

Figure. Electrophoretical spectra of maize androgenic regenerant And44(A), its donor hybrid H99xWf9(B), lines H99(C) and Wf9(D)

The spectrum of the pollen regenerant was close, but not identical to lines H99 and Wf9. The upper and middle part of the spectrum (high-molecular proteins) of the regenerant was similar to line H99, there were only differences in the intensity of some lines (2,7). The lower part of the regenerant spectrum had a resemblance with the parental line Wf9. Besides that, in the zone of minor components the regenerant had a line which was present neither in paternal nor in maternal lines (8).

The results of the morphobiological analysis of progeny of pollen regenerant and lines H99 and Wf9 are represented in the table.

Table. Morphobiological characteristics of the line of pollen origin And44 and the parental lines of the donor hybrid, H99 and Wf9.
 

Genotype	Plant height		Number of days from seedlings to tassel flowering		Number of days from seedlings to ear flowering
	Average, cm	Coefficient of variation, %	Average, days	Coefficient of variation, %	Average, days	Coefficient of variation,%
And44	132.9	6.55	75.1	4.13	76.4	5.66
H99	112.4	8.91	73.8	5.53	73.2	4.03
Wf9	162.7	5.99	70.2	5.45	70.8	6.62

Morphobiological analysis showed that the progeny of And44 had intermediate position between H99 and Wf9 in plant height. As for number of days from seedlings to tassel and ear flowerings it surpassed both of the lines. The coefficient of variation of And44, which may serve as indexes of the evening of a line, for different characteristics were on the level of the inbreds H99 and Wf9.

The evening and the identity of electrophoretical spectra of storage proteins (zeins) obtained from the first and the second self-pollinations of And44 may give evidence of the homozygosis of the androgenic regenerant. The data of electrophoretical analysis are well conformed with the results of the morphobiological estimation. The electrophoretical spectrum of the pollen regenerant differs from the spectra of maternal and paternal lines and donor hybrid and testifies to the obtaining of a new line.
 
 

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