In vitro haploid plant production of Zea mays L.

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Date
2012
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Birsa Agricultural University, Kanke, Ranchi, Jharkhand
Abstract
Maize (Zea mays L.) is the most important grain crop in several countries of the world. Due to its increasing importance to the world’s economy, it is essential that the maize germplasm base be maintained and enhanced for development of outstanding maize varieties. The development of homozygous lines by conventional methods is rather the time consuming procedure, for which haploid techniques can be utilized to increase the efficiency of breeding. Anther/pollen culture was the first promising technique to produce DH plants of microspore origin at satisfactory frequency. Much effort has been made to improve the in vitro responsiveness of various maize genotypes and optimize the culture conditions. However, genotype dependence is still one of the main limiting factors in the anther culture of maize, so another alternative method is to obtain haploid or DH plants from ovary or ovule culture. In the present study, it was tried that an efficient in vitro method for haploid plant regeneration via anthers/pollen and ovary culture of Z. mays (variety Suwan) be developed but the high genotype dependence and the low response rate limited haploid regeneration. The highest recorded decontamination percentage during surface sterilization of Z. mays (variety Suwan) explants were achieved by treating anthers with 0.05% of HgCl2 for 5 minutes and ovaries with 0.05% HgCl2 for 5 minutes. Absolutely no response was observed in anthers either in MS and N6 basal media or supplemented with casein hydrolysate. However, swelling of anthers was observed in MS supplemented with 2.0 mg/l 2, 4-D, N6 with 0.5 g/l charcoal and N6 with 2.0 mg/l 2,4-D + 1.0 mg/l Kn. While protrusion of filament like structure was observed in N6 supplemented with 2.0 mg/l and 6.0 mg/l 2,4-D, MS with 2.0 mg/l and 6.0 mg/l 2,4-D and in a combination of 2,4-D and Kn (2.0 mg/l 2,4-D + 2.0 mg/l Kn).While spikelets inoculated in MS supplemented with 2.0 mg/l 2,4-D, showed sprouting releasing the anthers. So far the microspores are concerned, no response was observed in all the combinations of media tried . On the contrary, ovary showed some response in different media . Colour change from cream to green was observed in N6 supplemented with 2.0 mg/l and 3.0 mg/l 2,4-D and MS with 2.0 mg/l and 6.0 mg/l 2,4-D . Sprouting of ovary was observed in MS basal liquid with 2,4-D and Kn combination, 2.0 mg/l each . 4 However, callusing was induced in the solid media with same combination as well as concentration of phytohormones.
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In vitro haploid plant production of Zea mays L.
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