scholarly journals Live-cell imaging of early events following pollen perception in self-incompatible Arabidopsis thaliana

2019 ◽  
Author(s):  
Frédérique Rozier ◽  
Lucie Riglet ◽  
Chie Kodera ◽  
Vincent Bayle ◽  
Eléonore Durand ◽  
...  

AbstractEarly events occurring at the surface of the female organ are critical for plant reproduction, especially in species with a dry stigma. Following landing on the stigmatic papilla cells, the pollen hydrates and germinates a tube, which penetrates the cell wall and grows towards the ovules to convey the male gametes to the embryo sac. In self-incompatible (SI) species within the Brassicaceae, these processes are blocked when the stigma encounters an incompatible pollen. Here, based on the generation of SI-Arabidopsis lines and by setting up a live imaging system, we showed that control of pollen hydration has a central role in pollen selectivity. The faster pollen pumps water from the papilla during an initial period of 10 minutes, the faster it germinates. Furthermore, we found that the SI barriers act to block the proper hydration of incompatible pollen and when hydration is promoted by high humidity, an additional control prevents pollen tube penetration into the stigmatic wall. In papilla cells, actin bundles focalize at the contact site with the compatible pollen but not with the incompatible one, raising the possibility that stigmatic cells react to the mechanical pressure applied by the invading growing tube.HighlightA live imaging system coupled with self-incompatible Arabidopsis lines highlight the role of stigmatic cells in controlling pollen hydration and in reacting to pollen tube intrusion by remodeling actin cytoskeleton.

2020 ◽  
Vol 71 (9) ◽  
pp. 2513-2526 ◽  
Author(s):  
Frédérique Rozier ◽  
Lucie Riglet ◽  
Chie Kodera ◽  
Vincent Bayle ◽  
Eléonore Durand ◽  
...  

Abstract Early events occurring at the surface of the female organ are critical for plant reproduction, especially in species with a dry stigma. After landing on the stigmatic papilla cells, the pollen hydrates and germinates a tube, which penetrates the cell wall and grows towards the ovules to convey the male gametes to the embryo sac. In self-incompatible species within the Brassicaceae, these processes are blocked when the stigma encounters an incompatible pollen. Based on the generation of self-incompatible Arabidopsis lines and by setting up a live imaging system, we showed that control of pollen hydration has a central role in pollen selectivity. The faster the pollen pumps water from the papilla during an initial period of 10 min, the faster it germinates. Furthermore, we found that the self-incompatibility barriers act to block the proper hydration of incompatible pollen and, when hydration is promoted by high humidity, an additional control prevents pollen tube penetration into the stigmatic wall. In papilla cells, actin bundles focalize at the contact site with the compatible pollen but not with the incompatible pollen, raising the possibility that stigmatic cells react to the mechanical pressure applied by the invading growing tube.


1973 ◽  
Vol 12 (2) ◽  
pp. 403-419 ◽  
Author(s):  
D. DE NETTANCOURT ◽  
M. DEVREUX ◽  
A. BOZZINI ◽  
M. CRESTI ◽  
E. PACINI ◽  
...  

The experimental results obtained show that the tip of the incompatible pollen tube bursts open after the outer-wall has considerably expanded in the intercellular spaces of the conducting tissue and the inner-wall has disappeared and numerous particles have accumulated in the tube cytoplasm. These particles, which measure approximately 0.2 µm in diameter and give a weak reaction to the test of Thiéry, differ in many respects from the vesicles normally present in compatible pollen tubes growing through the style; they appear to resemble, in some cases, the spheres which are discharged by the compatible pollen tubes after they have reached the embryo-sac. It is considered that these observations support the current belief that the tube wall is the site of action for the incompatibility proteins and suggest that self-incompatibility is not a passive process resulting from lack of growth stimulation but an active event which leads to the destruction of the incompatible pollen tubes. The degradation mechanism involved appears similar to the one which enables the compatible pollen tube to release its contents in the degenerated synergid and presents some analogies with the lytic process taking place in virus-infected cells. The general hypothesis is presented that the particles observed in the cytoplasm of self-incompatible pollen tubes consist of a mixture of incompatibility proteins and of basic constituents of the tube wall.


Author(s):  
Hyun Kyung Lee ◽  
Daphne R Goring

Abstract In flowering plants, cell–cell communication between the compatible pollen grain/growing pollen tube and the pistil is an essential component for successful sexual reproduction. In Arabidopsis thaliana, the later stages of this dialogue are mediated by several peptide ligands and receptors that guide pollen tubes to the ovules for the release of sperm cells. Despite a detailed understanding of these processes, a key gap remains regarding the nature of the regulators that function at the earlier stages which are essential steps leading to fertilization. Here, we report on new functions for A. thaliana Receptor-Like Kinase (RLK) genes belonging to the LRR-II and LRR-VIII-2 RLK subgroups in the female reproductive tract to regulate compatible pollen hydration and the early stages of pollen tube growth. Mutant pistils for the A. thaliana RKF1 gene cluster were observed to support reduced wild-type pollen hydration and, when combined with the SERK1 and SERK3/BAK1 mutations, reduced pollen tube travel distances occurred. As these mutant pistils displayed a wild-type morphology, we propose that the observed altered compatible pollen responses result from an impaired pollen–pistil dialogue at these early stages.


Zygote ◽  
1993 ◽  
Vol 1 (2) ◽  
pp. 143-154 ◽  
Author(s):  
Bing-Quan Huang ◽  
Elisabeth S. Pierson ◽  
Scott D. Russell ◽  
Antonio Tiezzi ◽  
Mauro Cresti

The cytoskeletal organisation of the isolated embryo sac and egg cells of Plumbago zeylanica was examined before, during and after pollen tube penetration into the embryo sac to determine the potential involvement of microtubules and actin filaments in fertilisation. Material was singly and triply stained using Hoechst 33258 to localise DNA, fluorescein isothiocyanate (FITC)-labelled anti- α-tubulin to detect microtubules and rhodamine-phalloidin to visualise F-actin. Microtubules in the unfertilised egg cell are longitudinally aligned in the micropylar and mid-lateral areas, aggregating into bundles near the filiform apparatus. In the perinuclear cytoplasm of the egg cell, microtubules become more or less randomly aligned. F-actin bundles form a longitudinally aligned mesh in the chalazal cytoplasm of the egg cell. In the central cell, microtubules and F-actin are distributed along transvacuolar strands and are also evident in the perinuclear region and at the periphery of the cell. During pollen tube penetration, sparse microtubule bundles near the pathway of the pollen tube may form an apparent microtubular ‘conduit’ surrounding the male gametes at the delivery site. Actin aggregates become organised near the pathway of the pollen tube and at the delivery site of the sperm cells. Subsequently, actin aggregates form a ‘corona’ structure in the intercellular region between the egg and central cell where gametic fusion occurs. The corona may have a role in maintaining the close proximity of the egg and central cell and helping the two sperm cells move and bind to their target cells. The cytoskeleton may also be involved in causing the two nuclei of the egg and central cell to approach one another at the site of gametic fusion and transporting the two sperm nuclei into alignment with their respective female nucleus. The cytoskeleton is reorganised during early embryogenesis.


2000 ◽  
Vol 6 (S2) ◽  
pp. 696-697
Author(s):  
Hua Zhu ◽  
Scott D. Russell

The embryo sac (ES) of Plumbago zeylanica, unlike most other flowering plants, lacks synergidsthe cell type that usually receives the pollen tube and the male gametes. Normally, synergids store copious amounts of calcium, estimated to exceed 15% by weight; this is believed to attract pollen tubes, which penetrate the ES, and may trigger the release of the sperm cells within one of the two synergids. If high concentrations of calcium are truly required for fertilization, the ES of Plumbago should also contain significant quantities. Synergids in normal flowering plants are preprogrammed for cell death, receiving the contents of the pollen tube directly in their cells, whereas synergid-lacking angiosperms apparently do not have such an apoptotic cell in the ES. Potassium pyroantimonate labeling was used to localize principally loosely-bound calcium, because it is a relatively mobile form of the Ca2+ ion that is available for redistribution during fertilization events.


1982 ◽  
Vol 60 (11) ◽  
pp. 2219-2230 ◽  
Author(s):  
Scott D. Russell

The ultrastructural organization of the megagametophyte of Plumbago zeylanica, which lacks synergids, was examined in chemically and physically fixed ovules after entry of the pollen tube. Similar to angiosperms with conventionally organized megagametophytes, the pollen tube enters the ovule through a micropyle, formed by the inner integument, and approaches the female gametophyte by growing between nucellar cells. Unlike other described female gametophytes, however, continued pollen tube growth results in direct penetration of the base of the egg through cell wall projections forming a filiform apparatus and is completed between the egg and central cell without disrupting either of these cells' plasma membranes. A terminal pollen tube aperture forms when the pollen tube reaches an area of strong curvature near the summit of the egg; this results in the release of two sperm cells, the vegetative nucleus, and a limited amount of pollen cytoplasm. The formerly continuous chalazal egg cell wall is locally disrupted near the tip of the pollen tube and apparently is thus modified for reception of male gametes. Discharged pollen cytoplasm rapidly degenerates between the egg and central cell, but unlike pollen tube discharge in conventionally organized megagametophytes, it is unassociated with the degeneraton of any receptor cell within the female gametophyte. Sperm nuclei are transmitted, one to the egg and the other to the central cell, to effect double fertilization by nuclear fusion with their respective female reproductive nuclei. The vegetative nucleus and discharged pollen cytoplasm degenerate between the developing embryo and endosperm during early embryogenesis. The emerging concept that the egg of Plumbago possesses combined egg and synergid functions is supported by the present study and suggests that the megagametophyte of this plant displays a highly specialized egg apparatus composed exclusively of a single, modified egg cell.


2020 ◽  
Vol 64 (1-2-3) ◽  
pp. 7-19
Author(s):  
Kundaranahalli R. Shivanna ◽  
Rajesh Tandon

Professor Panchanan Maheshwari served as Professor and Head of the Department of Botany, University of Delhi, from 1950 to 1966 and built an internationally reputed School of integrated plant embryology. Studies carried out during and after Maheshwari’s period from this School have enormously advanced our knowledge of the structural, developmental and functional aspects of embryological processes. This review covers studies carried out at the Delhi School on the developmental biology of dispersed pollen grains which operate from pollen dispersal from the anthers until pollen tubes discharge the male gametes in the embryo sac for fertilization. These events include pollen viability and vigour, pollen germination and pollen tube growth, structural details of the pistil relevant to pollen function, pollination and pollen-pistil interaction.


2021 ◽  
Vol 22 (5) ◽  
pp. 2603
Author(s):  
Ana Marta Pereira ◽  
Diana Moreira ◽  
Sílvia Coimbra ◽  
Simona Masiero

Angiosperm reproduction relies on the precise growth of the pollen tube through different pistil tissues carrying two sperm cells into the ovules’ embryo sac, where they fuse with the egg and the central cell to accomplish double fertilization and ultimately initiate seed development. A network of intrinsic and tightly regulated communication and signaling cascades, which mediate continuous interactions between the pollen tube and the sporophytic and gametophytic female tissues, ensures the fast and meticulous growth of pollen tubes along the pistil, until it reaches the ovule embryo sac. Most of the pollen tube growth occurs in a specialized tissue—the transmitting tract—connecting the stigma, the style, and the ovary. This tissue is composed of highly secretory cells responsible for producing an extensive extracellular matrix. This multifaceted matrix is proposed to support and provide nutrition and adhesion for pollen tube growth and guidance. Insights pertaining to the mechanisms that underlie these processes remain sparse due to the difficulty of accessing and manipulating the female sporophytic tissues enclosed in the pistil. Here, we summarize the current knowledge on this key step of reproduction in flowering plants with special emphasis on the female transmitting tract tissue.


1986 ◽  
Vol 34 (4) ◽  
pp. 413 ◽  
Author(s):  
EG Williams ◽  
V Kaul ◽  
JL Rouse ◽  
BF Palser

Frequent overgrowths of pollen tubes within the embryo sac are characteristic of a number of interspecific crosses in the genus Rhododendron (Ericaceae). The combined techniques of sectioning, squashing and whole-ovule clearing have confirmed that in ovules showing this phenomenon the pollen tube fails to terminate growth and release sperms on entry into a synergid; instead it continues to grow beyond the synergid and egg cell, often filling the main body of the embryo sac with a coiled and distorted mass. Such ovules fail to develop further. The occurrence and possible causes of this error syndrome are discussed.


1982 ◽  
Vol 214 (1195) ◽  
pp. 273-283 ◽  

Cratoxylum formosum shows all the classical features of a distylic species. The two types are: long-styled plants with short stamens and small pollen grains and short-styled plants with long stamens and large pollen grains. Compatible pollinations are only between the two types; incompatible pollen tubes are inhibited in the style. A significant morphological feature distinguishes Cratoxylum from distylic plants in other families. Instead of having a small number of anthers making well separated narrow discs in the two types, Cratoxylum has many anthers (144) and they are arranged on staminal bundles that produce long cylinders of anthers that partially occupy similar height zones in the two types of flower. A novel method of separation of the two height zones is achieved by the bending of the stamens of the long-styled type when the flower opens, which converts the cylinder to a narrow disc of anthers at the same height as the ‘short’ stigma. The bending coincides with anther dehiscence and is slightly later than the first daily insect visitation. The anthers return to the upright position later in the day, when the pollination is complete. There was a 20-fold difference between the amounts of pollen deposited on the two types of stigmas. The ‘long’ stigmas received 1200 pollen grains per flower, in the ratio of 46 ‘long’ to 54 ‘short’, which is close to the ratio of two types of pollen produced in the population. This random deposition of pollen on ‘long’ stigmas is, however, more than adequate for the 36 seeds produced per flower. ‘Short’ stigmas received only 64 pollen grains per flower, in the ratio of 90 ‘long’ to 10 ‘short’, and several flowers had below the critical level of 36 compatible pollen grains for full seed production. Pollen loads of the pollinating bee, Apis javana , consisted of ‘long’ and ‘short’ pollen on the thorax in the ratio found on the ‘long’ stigma, and on the head of the bee in a ratio close to the 9:1 found on the ‘short ’ stigma. The corbicular loads reflected accurately the pollen of the tree in which the bee was caught. For Cratoxylum the accurate positioning of the anthers of the long-styled plant in relation to the visiting bees head was an important evolutionary step in the effective pollination of the short-styled form, which, at least in this species, is one critical and highly selected feature of the system.


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