Pollen Viability and Microspore Culture in Three Broccoli Cultivars (Brassica oleracea L. var. italica Plenck)

Broccoli is a high value vegetable crop in Indonesia, however production is low due to limited number of suitable cultivars, so, breeding hybrid broccoli for warm climate is important. The first step in hybridization is providing homozygote parent plants which can be done efficiently via microspore culture. The objectives of this study were to determine 1) bud size that produce uninucleate microspore stage appropriate for culture; 2) pollen viability, 3) microspore development, in three broccoli cultivars (‘BL 10001’, ‘Royal Green’ and ‘Green Magic’). Various bud size (1 – 5 cm) was squashed and observed microscopically to determine bud size containing uninucleate microspore. Pollen viability was determined by IKI staining and pollen germination method. Chromosome number was counted on root tips using squash method with aceto-orcein stain. Various heat treatment schemes were conducted to induce microspreo development. Result showed uninucleate microspore derived from 2-3 mm and 3-4 mm bud length of ‘BL-10001’ and ‘Royal Green’ was responsive for microspore development in culture. Pollen viability varied among cultivars, 78-87% on IKI method and 15-16% on germination test. Microspore culture showed different embryogenesis response; pollen-like structure was produced by ‘BL 10001’.

SELECTION OF SOYBEAN CULTIVARS: PREPARATION THE PLANT BREEDING WITH MICROSPORE CULTURE METHOD

Microspore culture method can be used as plant breeding program. The preparation of cultivars selection is an important step. The research starts with cultivated five cultivars of soybean i.e: Argomulyo, Grobogan, Wilis, Anjasmoro and Black Malika. The appearence of soybean plant was observed until flowering. Selection of plants based on: sum of flower bud every plant, anther midline, total and diameter of microspore every flower bud was measured by the ‘Optilab’ software. The development of microspore done with grouping of flower bud according long 2.02.5 mm, 2.6-3.0 mm, 3.1- 3.6 mm and 3.7-4.1 mm for chooses flower bud with the most late uninucleate microspore stadium. Result of the research shows that long of flower bud 2.6-3.6 mm contain 1847-2010 late uninucleate microspores, diameter 20 µm for 5 cultivars can be used for material of microspore culture. Anjasmoro cultivar, tall of plant gain 68 cm, sum of rame 7-9, anther midline 354.67±59.67 µm, number of microspores each flower bud 2003±216. Result of responsive qulity test with anther incubation on 340C temperature for 4 days represent the most of total viable microspore, 3.371±45 on Anjasmoro cultivar. Plant breeding by Anjasmoro cultivar is the most appropriate for microspore culture treatment.

PROPORSI MIKROSPORA UNINUKLEAT PADA EMPAT KLON TEBU (Saccharum spp.)

An experiment to study the two different morphological characters of four clones of sugarcane (Saccharum spp.) panicles containing more than 50 percent of uninucleate microspore development was conducted in Tissue Culture Laboratory at Biology Faculty, Gadjah Mada University, Yogyakarta, since March until May 2006. Morphological characters of both kinds of panicles i.e. unsheated- and sheated flowers from sheat flag leaf were observed. Mean, percentage, and standard deviation from the mean value of the six different stages of microspore development e.g. tetrad, early- and late-uninucleate, early- and late-binucleate, and multinucleate or pollen grains were statistically used in this calculation. All data percentages were analyzed by variance analysis through General Linier Model Procedure, and comparisons between means of the uninucleate microspore development based on the two different morphological characters of four clones was calculated by Least Significance Difference method. Comparisons between the two different panicles characteristics in accordance with the proportion of the uninucleate microspore development, however, were analyzed by T-student procedure. All calculations were done by using SAS program of computer statistics package. Result of the research showed that: (1) the unsheated panicles were contained less than 50 percent of uninucleate (early- and late-uninucleate) microspore development; (2) the sheated panicles tend to be in high proportion of early- and late-uninucleate microspore development, and multinucleate or pollen grains, and (3) the more away of spikelets or anthers positioned in the panicle or sub-panicle, the more number or percentage of uninucleate microspores development were tend to be gradually decreased.