Hey, what's up everyone? Welcome to my YouTube channel LifeSense with KMD. Today I'm going to explain about Anther and Poland culture. Now, first is what is an Anther. Anther is a part of Stamen containing Poland grains. Coming to the history of Anther or Poland grains, two-leg in 1953 first obtained the major Poland grains of Ginkgo-Byloba, which is a gymnosperm, can be induced to proliferate in culture to form haploid gallows. Goha and Maheshwari in 1964 first reported the direct development of embryos from microspores of the dura in Oksia. By the culture of exized Anther, Borgen and Hnist in 1967 obtained complete haploid plantlet from Anther culture of Nicotiana Tobakum. Now, what is anther culture? Culturing of Anther's obtained from unopened flower bird in the nutrient medium under aseptic condition. Calus tissue or embryoid from Anther that give rise to a haploid plantlet either through organogenesis or embryogenesis. Now, what is a Poland culture? Poland or microspore culture is an inviterate technique by which Poland grains preferably at the unineucleated stage or squeezed out heseptically from the intact anther and then cultured on nutrient medium. The microspore developed into a haploid embryo or calus tissue that give rise to haploid plantlet by embryogenesis or organogenesis. Androgenesis is the inviterate development of haploid plant originated from 2T potent Poland grain through a series of cell division and differentiation. It is of 2 type direct and direct androgenesis that is the microspore behave like a zygote and undergo change to form a embryoid which ultimately give rise to a plantlet. Another one is the second one is indirect androgenesis. Here the microspore divide repeatedly to form a calus tissue which differentiate into haploid plantlets. Principle of anther and Poland culture. The production of haploid plant is to exploit and to T-potency of microspores. In this process the normal development and function of the Poland cell to become a male gamut is stalked and is diverted forcefully to a new metabolic pathway for vegetative cell division. Methods of anther culture. Collection of unopened flower bird. Surface sterilized with 70% ethanol. Anther is excised from flower bird and kept separately. Anther is first in anther in first mytotic division is selected by acetocormide test. Inoculated in the medium containing glutamine L-serene inocetol. Incubate the culture at 25 degrees centigrade for 15 days. Here anther grows into embryoid. Embryoid transfer to rooting medium under 3000 luxe illumination. After 3 to 5 weeks the embryoids become plantlets for acclimatization transfer into the greenhouse. In this picture how anther culture will be done will be explained. First we will select the plant of our interest and then from that flower bird unopened flower bird will be taken. Later on it will be surface sterilized with 70% ethanol and sodium hypochloride solution. This bird was next sterilized with distilled water. Then split flower bird and take cut on the stamens. Later on anther in first mytotic division is selected by acetocormide test. Later the anther like discard the filament and just take the anther. Now the anther in the first mytotic division is selected by acetocormide test. Then it will be inoculated into a medium containing glutamine and L-serene and inocetol. Then we have to incubate this culture at 24 to 26 degrees centigrade in dark for 3 to 4 weeks. Later on when either this anther will be converted into direct undergo a direct androgenesis pathway to form a plantlet or it could undergo callus form. Callus formation later on organogenesis and then it will form a plantlet which is a haploid regenerated plantlet will be formed. Methods of pollen culture and they are collected from flower birds and pollen grains are isolated and about 50 anther are placed in a small sterile beaker containing 20 ml of liquid basal medium. It could be either MS or white or next medium. Anther is then pressed against the side of the beaker with a sterile glass rod to squeeze out the pollen. The homogenized anther are then filtered through nylon sleeve to remove the anther tissue debris. Filter or pollen suspension is then centrifused at low speed for 5 minutes. The supernatant containing fine debris is discarded and pellet of pollen is suspended in fresh liquid medium and washed twice by repeated centrifugation and resuspension in the fresh liquid medium. 2.5 ml of pollen suspension usually 10 to the power of 3 to 10 to the power of 4 per ml is pipetted off and is spread in 5 centimeter predatory dish. Pollen are best grown in liquid medium but if necessary they can be grown by plating very soft agar added medium. Petri dish are incubated at 20 to 30 degrees centigrade under low intensity of white cool light. Young embryoid can be observed after 30 days the embryoids ultimately give rise to a haploid plant let. Influence of anther culture or pollen culture, genotype of donor plant. Genotype of donor plant play a significant role in determining the frequency of pollen production. For example, Hordeum of each genotype differ with respect to androgenic response in anther culture. Stages of microspores and thus are more productive when cultured in unineucleated microspores stage. For example, barley wheat rice etc. Culture medium, anther pollen culture, medium requirement, berry with genotype and age of the anther as well as condition under which donor plant are grown. Incorporation of activated charcoal into the medium has stimulated the induction of androgenesis. Iron in the medium play a very important role for the induction of haploids. Portatoextract, coconut milk, growth regulators like oxen cytokine lins are used for anther and pollen culture. Temperature, temperature enhance the induction frequency of micro-spor androgenesis. The low temperature treatment to anther or flurbered hennands the haploid formation. Low temperature affect the number of factors such as dissolution of microtubules lowering the absurcesic acid, maintenance of high ratio of viable protein capable of embryogenesis. Physiological status of donor plant such as water stress, nitrogen requirement and age of donor plant highly affect the pollen embryogenesis. Plant, start of nitrogen may give more responsive anther compared to those that are well fed with nitrogen as fertilizer. Advantage of anther culture, simple, time less time consuming and responsive. Disadvantages, requires skill to remove anther's without causing damage, not much successful in case of serial crops, risk of chimeras and chalice formation from anther walls. Application of anther and pollen culture, it is used for mutation study, plant breeding and crop improvement and it is used for secondary metabolites. Even haploid are used in molecular biology and genetic engineering, haploid tissue of harabodopsis and lycopasicum have been used for the transfer and expression of three genes from esterysia coli. Thanks for watching my video, please subscribe to my channel by clicking on subscription button. Subscription doesn't cost you any money to get a notification click on a bell icon, do like and share this video with friends and family. If you have any kind of feedback do share it on a comment box. Thank you.