By Christophe Clement, Ettore Pacini, Jean-Claude Audran
Pollen, the plant constitution most generally utilized by people, is a key constitution in plant copy giving upward thrust to culmination and seeds. additionally, the biotechnological use of pollen is of serious value for plant breeders because it permits to procure types with higher usage and yield. within the first half, the successive steps of pollen improvement within the anther from floral induction to pollen germination and fertilization are completely tested; the second one half is dedicated to pollen behaviour in vitro.
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The location of these genes was confirmed by segregation analysis using Southern blot analysis: each of the cDNA hybridised to a genomic DNA fragment which strictly segregates with the male plants. Interestingly, all 5 clones seem to have a homologue on the X chromosome. The sequence analysis of these clones revealed that they are all similar to EST sequences (Expressed ~equence lags) of Arabidopsis or rice. One of them has 2 calcium binding domains similar to those found in calmodulin. Another one has strong similarity to Rb-binding proteins found in mammals and we thereofre focused our work on this clone.
We also studied ubiquitin involvement during the differentiation of microgametophytes in male and female kiwifruit flowers. Microgametophyte development in male flowers was characterized by a modulation of protein ubiquitination. By contrast, a progressive decrease of free and conjugated ubiquitin occurred in that of female ones due to the altered metabolic activity of this microgametophyte which is programmed to degenerate. A large amount of peak volume (pi ~CI inlcnsi lyl J()()()) 3m . . - - - - - - - - - - - - - - .
Later on, a dramatic loss of ubiquitination occurred at days 29 and 35, when only two small peaks were detectable. Free ubiquitin levels changed strongly during microgametophyte differentiation: two opposite trends marked the developmental path in male and female flowers (Fig. 4). While a continuous increase of free ubiquitin occurred in the male until maturity, in the microgametophyte of female flowers the largest amount was detected at the tetrad stage, then it rapidly decreased. 4 Discussion Anther dehiscence is a complex process which starts when reproductive cells are at the tetrad stage (Goldberg et al.