Báo cáo lâm nghiệp: "Sexual reproduction in Populus I. Some physiological and biochemical events of the progamic phase"

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Sexual reproduction in Populus I. Some physiological and biochemical events of the progamic phase C. Dumas 2 2 Gaget M. Villar M. 1 INRA, 1 Station D’Amélioration des Arbres Forestiers, Ardon, F-45160 Olivet, and dAm6lioration Laboratoire 2 de Reconnaissance Cellulaire et Am6lioration des Plantes, INRA 23879, Université Lyon I, 43, bd 11-Novembre-1918, F-69622 Villeurbanne Cedex, France tions, silver nitrate staining and homogenates of Introduction pollinated stigma performed according to were the procedures ot’ Villar et al. (1988). Each of the 2 crosses was represented by 3 series of Hybridization in Populus breeding pro- stigmatic extract!., in accordance with the grams is limited by sexual incompatibility growth kinetics (0.6 and 20 h after pollination). barriers, whose cellular and molecular Protein patterns of these extracts were com- mechanisms are not yet known. In an pared on a single gel. Glycoprotein revelation (concanavalin A-binding proteins) on IEF poly- attempt to understand the nature of inter- acrylamide gel was adapted from Hawkes specific incompatibility in Populus, we (1982). j3-Ga!actosidase visualization was op- have explored the interactions between timized from the protocol of Singh and Knox male and female partners (pollen-pistil) in (1985). compatible and incompatible crosses P. nigra (female) x P. nigra (male) and P. nigra (female) x P. alba (male) (Villar, Results and Dis;cussion 1987). Kinetics of pollen tube growth (visualiza- tion using the AI3F method) have demon- Materials and Methods strated distinct behaviors of P nigra and P alba pollen tubes inside P nigra pistils (Fig. 1P alba pollen tubes exhibit a P. nigra and P. alba branches were obtained unique S-shaped growth curve and an from the INRA Forestry station in Or[6ans (France). Pollen was collected and stored in arrested growth near the stylodium. On closed vials at -18°C. Kinetics of pollen tube the other hand, P nigra pollen tubes ex- growth in vivo were studied in growth chambers hibit 2 growth phases, respectively, in the (20°C). Pollen tubes in the stylar tissues were stigmatic tissues and in the ovarian cavity. visualized using the aniline blue fluorescence P, nigra and P alba curves diverge 5 h method (ABF method, Dumas and Knox, 1983). after pollination (20°C), corresponding Isoelectric focusing (IEF) of pollinic and stig- matic proteins, gel preparation, focusing condi- precisely to the deposition of the first
observed, Differences plug inside pollen tubes. This callose glycoproteins. were distinct type of to the divergence could likely be related to a according cross: one in the compat- increases change in the physiology of the compat- only glycoprotein ible cross. Moreover, f3-Galactosidase ac- ible pollen tube, shifting from an auto- tivities were revealed with a similar trophic to a heterotropic type of nutrition. electrophoretic technique in pollinated Biochemical studies focused on pollinic stigma. This pollinic enzyme could play a and stigmatic proteins, known to be in- role in heterotropic pollen tube nutrition volved in male-female interactions (Singh and Knox, 1985). An increase of its (Gaude and Dumas, 1987). After silver activity (one isoenzyme of isoelectric point nitrate staining, qualitative and quantita- about pH 4.2) from 6-20 h after pollination tive differences could be observed, related was detected only in the compatible cross. to the presence of P nigra and P alba pol- len tubes inside stigmatic tissues. How- ever, increasing protein bands were Conclusion detectable, 0-20 h after pollination, only in compatible pollinated stigmas. The conca- in P. The compatible progamic phase navalin A-peroxidase reaction allowed the to the nigra, i.e., pollen tube growth up visualization of 15 stigmatic and pollinic
sac, could be related to pollinic Gaget M. (1988) incompatibility intersp6cifique embryo chez Populus: effet Mentor. Ph.D. Thesis, Uni- enzymes involved in pollen tube metabo- versite Lyon I, France lism, such as ¡3-galactosidase. Its activity Gaude T. & Dumas C. (1987) Molecular and could be the final result of a series of inter- cellular events of self-incompatibility. Int. Rev. actions started by initial pollen-stigma Cytol. 107,333-366 communications. This dialogue probably Hawkes R. (1982) Identification of concanavalin implicates protein compounds, detected in A-binding proteins after sodium dodecyl sul- pollen and pollen tube diffusates in vitro fate-gel electrophoresis and protein blotting. AnaL Biochem. 4 23, 143-146 (Villar, 1987; Gaget, 1988). Singh K. and Knox R.B. (1985) P-Galactosidase of Lilium pollen. Phytochemistry 24, 1639-1643 Villar M. (1987) Incompatibilit6 intersp6cifique chez Populus: approches physiologique et References biochimique. Phi.D. Thesis, Universit6 Lyon I, France Dumas C. & Knox R.B. (1983) Callose and Villar M., Gaget M. & Dumas C. (1988) Micro- determination of pistil viability and incompatibili- isoelectric focusing of proteins from single stig- ty. Theor. AppL Genet. 67, 1-10 ma in Populus. Can. J. For. 18, 1261-1264 0