Pollen or microspore culture is an in vitro technique by which the pollen grains are squeezed out asep­tically from the intact anther and then cultured on nutrient medium. the microspores, without producing male gametes, develop into haploid embryoids or callus tissue that give rise to haploid plantlets by embryogenesis or orga­nogenesis. this helps to acquire the whole knowledge about anther and pollen culture.

1. ANTHER AND POLLEN
CULTURE
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2. WHAT IS ANTHER?
A part of stamen containing pollen.

3. WHAT IS POLLEN?
• A fertilizing powder discharge from flower’s anther.

4. HISTORY
• W.TULECKE(1953)
First observed that mature pollen grains of Ginkgo biloba
(gymnosperms) can be induced to proliferate in culture to form
haploid callus.
• S.GUHA AND S.C.MAHESHWARI (1964)
First reported the direct development of embryos from microspore of
Datura innoxia, by the culture excised anther.
• J.P.BOURGIN AND J.P.NITSCH (1967)
Obtained complete haploid plantlets from anther culture of nicotiana
tabacum.

5. WHAT IS ANTHER CULTURE?
• Anther culture is a technique by which the developing of anthers at a
aseptically from unopened flower bud and are cultured on a nutrient
medium.
• The microspores within the cultured anther develop into callus tissue
or embryoids that give rise to haploid plantlets either through
organogenesis or embryogenesis.

6. WHAT IS POLLEN CULTURE?
• Pollen or microspore culture is an in vitro technique by which the
pollen grains are squeezed out aseptically from the intact anther and
then cultured on nutrient medium.
• the microspores, without producing male gametes, develop into
haploid embryoids or callus tissue that give rise to haploid plantlets
by embryogenesis or organogenesis.

8. WHAT IS ANDROGENESIS?
• Androgenesis is the in vitro development of haploid plants
originating from totipotent pollen grains through a series of
cell division and differentiation

9. THERE ARE TWO MODES OF ANDROGENESIS:
(i) Direct androgenesis:
• In this type, microspore behaves like a zygote and undergoes change
to form embryoid which ultimately gives rise to a plantlet.
(ii) Indirect androgenesis:
• In contrast to the direct androgenesis, the microspore, instead of
undergoing embryogenesis, divide, repeatedly to form a callus tissue
which differentiates into haploid plantlets.

10. STEPS INVOLVED IN ANTHER AND POLLEN
CULTURE
• STEP1:
 Collect flower bud (normally 3-4months old plant)
 measures about 17-22mm in length.
 reject all flower buds which are beginning to open.
• STEP2:
 Transfer the selected flower bud to the laminar air flow chamber.
 flowers are allowed to surface sterilization with 70% ethanol of 10seconds followed
by 10minutes in 2%sodium hypochlorite.
 then wash 3times with sterile distilled water and transfer to sterile petridish.

11. • STEP3:
To remove anther,
slit the slide of the bud with a sharp instrument and remove them.
with the help of foreceps, place the anthers with filaments to another
petridish.
filaments are cut gently and ensure that anther are not injured.
• STEP4:
anthers are placed on MS medium or Nitsch and Nitsch medium.
• STEP5:
 the culture kept initially in dark.
 after 3-4weeks the anthers are normally undergo pollen embryogenesis and
haploid plantlets appear from the culture anther.
 In somecases, anther may from callus tissue which can induce to differentiated
into haploid plants.

12. • STEP6:
 The cultures are incubated at 24-28degree centigrade in a 14hrs
daylight at 2000lux.
• STEP7:
 Approximately 50mm tall plantlets are freed from agar gently.
 Washed with running tapwater and transfer to small
pot(autoclaved) and cover with a glass beaker for prevention.
 after 1week, remove glass beakers and transfer to natural
environment and allow to adaption will takes place and it lead to
mature and finally flowers are seen.

13. HOMOZYGOUS PLANTS ARE ALWAYS
STERILE. WHY?
•(n) Univalent chromosomes are
always non-viable.

14. CONVERSION OF STERILE INTO FERTILE
• COLCHINE TREATMENT(indole alkaloid)
chromosome duplication will be induced by colchine treatment
4%.
colchine will inhibit the formation of spindle fibres (metaphase)it
leads to polyploidy.
• ENDOMITOSIS
Endomitosis is one of the processes leading to endopolyploidy, in
which the stages of mitosis (prophase, metaphase, anaphase) take place
inside the nuclear membrane and without spindle formation.

15. ADVATAGES
• The stage of androgenesis can be observed starting from single cell.
• Pollen may be directly transformed into an embryoid. So it is very
suitable for understanding biochemistry and physiology of
androgenesis.
• Higher yield of haploid plants per anther could be expected in pollen
culture.
• Overcrowding of pollen grain in anther is eliminated and isolated
pollen grains are equally exposed to nutrient medium.

16. DISADVANTAGE
• the plants or embryos or calli not only originate from microspore or
pollen but may also originate from various other parts of anther
lobes (anther wall, nucellus, tapetum) which are diploid tissues.

17. APPLICATIONS
• Haploid production – Major application of pollen / anther culture is production of
haploid plants. It takes much time to produce haploid plants by conventional
breeding methods (many generations of inbreeding or backcrossing). As pollens are
haploid, plants developed from these are homozygous (haploids).
• Protoplast isolation – Used for protoplast isolation as single pollens (unicellular)
are available.
• Transformation – Used in transgenic plant formation, it can be done with less
time consumption.
• Crop improvement – In-vitro anther culture is used for improvement in
vegetable and cereal crops e.g. asparagus, sweet pepper, watermelon, cabbage,
broccoli, wheat etc.

18. CONCLUSION
• THERE ARE PROS AND CONS ARE THERE IN ANTHER AND POLLEN
CULTURE.
• BUT IT IS BENEFICIAL TO HUMANRACE OF NOW-A-DAYS.

19. REFERENCES
• ANTHER AND POLLEN CULTURE TO PRODUCE HAPLOIDS: PROGRESS AND
APPLICATION FOR THE PLANT BREEDER1 K. C. Sink, Jr. and V. Padmanabhan
HortScience, Vo l.12(2), April 1977.
• https://www.plantcelltechnology.com/blog/production-of-haploids-from-microspore-
culture/
• Haploid production. (1996). Studies in Plant Science, 167–213.
• Farinha, Nelson. (2013). Pollen embryogenesis for rice plant production from selected
genotypes cultivated in the Mondego river valley.
• https://www.biologydiscussion.com/plant-tissues/pollen-culture/pollen-culture-meaning-
and-advantages-biotechnology/61368
• https://biologyreader.com/anther-culture.html
• https://naldc.nal.usda.gov/download/42088/PDF

20. DOUGHTS?!!

21. THANK YOU