Vegetable improvement via pollen grain and anther by tissue culture

introduction Plant tissue culture comprises a set of in vitro techniques, methods and strategies that are part of the group of technologies called plant biotechnology. Tissue culture has been exploited to create genetic variability from which crop plants can be improved, to improve the state of health of the planted material .Tissue-culture protocols are available for most crop species, although continued optimization is still required for many crops. Tissueculture techniques, in combination with molecular techniques, have been successfully used to incorporate specific traits through gene transfer. In vitro techniques for the culture of protoplasts, anthers, microspores, pollen grain, ovules and embryos have been used to create new genetic variation in the breeding lines, often via haploid production. Cell culture has also produced somaclonal and gametoclonal variants with crop-improvement potential. The culture of single cells and meristems can be effectively used to eradicate pathogens from planting material and thereby dramatically improve the yield of established cultivars. Large-scale micropropagation laboratories are providing millions of plants for the commercial ornamental market and the agricultural, clonallypropagated crop market. With selected laboratory material typically taking one or two decades to reach the commercial market through plant breeding, this technology can be expected to have an ever increasing impact on crop improvement as we approach the new millennium.

Objective: Iraqi researchers still use classic plant breeding methods in producing vegetable. This forms a serious constrain to increase productivity .Therefore new approaches in plant breeding should be applied to achieve this goal, especially. As these new approaches take short time, low cost and under controlled

condition. As well known, anthers and pollen grains have haploid (n chromosomes).By doubling them, and then following a procedure on these cells includes screening, selecting to determine the tolerated cells .Eventually, these cells differentiate to plants .These plants are transformed to open field to get their seeds for propagating it. This technique not available in Iraq. Because the Iraqi labs lack required equipments with no experience .The main objective of this study is to produce the vegetable like the tomato. Materials and methods 1-collection anthers form crop flowers at suitable stage of growth to determine the developmental stage of pollen grains. 2-sterilizing anthers and pollen grains with different concentrations of sodium hypo chloride for ten minutes and washed to remove the residual effects, of sterilized materials on pollen grain viability. 3-inducing callus by culturing anthers on media supplied with different concentrations of auxin, cytokianin,2,4-D,NAA and incubating them in dark for five weeks. 4-applying a sub-culture process to propagate induced callus. 5-screening and selecting the callus that cultured on the same media which gave a high rate of callus supplied with different concentrations of PEG. 6-diagnosing (LD50, LD100) to determine the best concentrations that the callus will be propagated and screed 7-doubling chromosomes from (n) to (2n) by colchicines 8-growning callus on differentiated media with different concentrations of auxin to promote the vegetative growth. 9-transfering the plant to the media supplied with different concentrations of auxin to encourage roots formation

10-Acclimitizing the rooted plantlets by transferring them to the green house 11-applying PCR test to determine the differences between the plant that derived from induced callus anther and their parents.

Constraints of conducting this study in Iraq 1- Specialized labs are not available 2- Lacks of some chemical materials from well known companies such as sigma. 3- Researchers have a little experience. 4- There are few controlled green houses to make screening.