Growth ofTrifolium pratense L. pollen tubes in compatible and incompatible styles of excised pistils

Summary The stigma and style portions of red clover pistils were culturedin vitro after cross- (genetically compatible) or self- (incompatible) pollination. Pollen tubes grew through styles in which they were compatible and, in some treatments, through styles which were incompatible.Boric acid, calcium nitrate and a carbohydrate in the medium, and moderate-to-cool temperatures generally enhanced pollen tube growth of compatible and incompatible matings. Several plant hormones at high concentrations inhibited pollen growth with compatible matings and did not retard the incompatibility mechanism. Application of high temperatures to the flower heads during the period of anthesis retarded the incompatibility mechanism. This temperature treatment affected the styles and not the pollen. In one of three experiments the application of relatively large amounts of pollen to the stigma rendered the incompatibility mechanism less effective.

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Zusammenfassung Narben und Griffel von Rotklee-Stempeln wurden nach Fremd- (genetisch kompatibel) oder Selbstbestäubung (inkompatibel) in vitro kultiviert. Die Pollenschläuche wuchsen durch die Griffel, mit denen sie kompatibel waren, und nach bestimmter Behandlung auch durch Griffel, mit denen sie inkompatibel waren. Ein Zusatz von Borsäure, Kalziumnitrat und einem Kohlenhydrat zum Nährmedium sowie mäßige bis kühle Temperaturen förderten im allgemeinen das Pollenschlauchwachstum der kompatiblen und inkompatiblen Kombinationen. Verschiedene Wuchsstoffe, die dem Kulturmedium zugesetzt wurden, hemmten bei höherer Konzentration das Pollenschlauchwachstum kompatibler Kombinationen, verzögerten auf der anderen Seite aber nicht den Inkompatibilitätsmechanismus. Letzterer Effekt trat ein, wenn die Blütenköpfe während der Anthesis hohen Temperaturen ausgesetzt wurden. Diese Temperaturbehandlung wirkte auf die Griffel, nicht auf den Pollen. In einem der drei Experimente führte eine Bestäubung der Narbe mit großen Pollenmengen dazu, den Inkompatibilitätsmechanismus weniger wirksam werden zu lassen.

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Cooperative research by Crops Research Division, Agricultural Research Service, U. S. Department of Agriculture and the Kentucky Agricultural Experiment Station. This paper (No. 68-3-23) is published with permission of the Director, Kentucky Agricultural Experiment Station.     

Pollen tube growth following compatible and incompatible intraspecific pollinations in Petunia hybrida

The observation that both compatible and incompatible pollen tubes grow at identical speeds on the stigma in many plants with gametophytically controlled self-incompatibility (SI) systems has, in Petunia, been extended to cover all other facets of pollen behaviour on this tissue. On entry into the stylar transmitting tissue both types of tubes accelerate, but the compatible achieve a higher terminal velocity than do the incompatible, which eventually slow and stop. Grafting experiments show that the top 1 mm of the stylar tissue can play an important rôle in determining the future development of the pollen tube. Following mixed pollinations, proportionally too many compatible pollen tubes reach the ovary than would be expected from the results of pure compatible and incompatible pollinations indicating that incompatible pollen in some way helps prime the style for growth of compatible pollen tubes. This data is considered in terms of recent structural studies of these tissues, and related to the pollination conditions pertaining to Petunia populations in the field.Abbreviation SI self-incompatibility     

Pollen tube behavior in yellow passion fruit following compatible and incompatible crosses

Pistil squashes were used to examine the growth of Passiflora edulis f. flavicarpa Deg pollen tubes in self-compatible and self-incompatible crosses. Compatible tubes typically showed a uniform layer of callose deposition in the walls and in small plugs spaced at regular intervals within the tube. Two sites of rejection were observed in incompatible crosses: the stigma and on some occasions the style, especially in anomalous crosses. In the style the inhibition of pollen-tube growth occurred in both the upper and middle parts of the transmitting tract. These findings are consistent with the hypothesis that suggests the presence of a gametophytic gene which acts in association with the sporophytic S-gene in P. edulis. Received: 12 October 1998 / Accepted: 2 December 1999     

Environmental Pollution: Leaf Cuticular Patterns in Trifolium pratense L.

Red clover populations collected from the polluted environmentsof Memphis, Nashville, and Knoxville (Tennessee) showed decreasedfloral production, low stomatal frequency, high trichome density,and longer trichomes. The results for plant population collectedfrom a less polluted, rural Reelfoot Lake area were quite theopposite. Subsidiary cell complex remained the same in all thepopulations, while stomatal size range showed little variation.     

Plant regeneration of natural tetraploid Trifolium Hum pratense L

The regeneration of natural tetraploid T. pratense, originated from Erzurum-Turkey, is reported in this study. This plant has low seed setting and hard seed problems due to polyploidy. Hypocotyl, cotyledon, apical meristems, epicotyl and young primary leaves were inoculated on MS and PC-L2 media containing different concentrations of BAP and NAA as growth regulators. The best shoot formation has been observed on explants initiated from apical meristem placed on PC-L2 medium that includes 2 mg dm-3 BAP and 1 mg dm-3 NAA. 94.4% of the shoots originated from calli were rooted on PC-L2 medium with 1 mg dm-3 NAA. In vitro organogénesis has been accomplished in the natural tetraploid T. pratense regenerated plants successively transferred to the field.     

Plant regeneration from hypocotyl and petiole callus of Trifolium pratense L.

Red clover (Trifolium pratense L.) seedlings were screened for the ability to regenerate plantlets from hypocotyl-derived callus tissue. Media sequences described by Beach and Smith (1979) and Collins and Phillips (1982) and a variation using media from both sequences were tested. Plantlets were regenerated from three out of 642 genotypes. In all three cases, callus was initiated on B5C medium and regeneration was accomplished on SPL medium. Attempts to regenerate plants from petiole-derived callus tissue have so far been successful only with regenerants of clone F49. Petiole callus from epicotyl-derived F49 plants proved to be non-regenerative. Pollen viability varied significantly among individuals regenerated from callus cultures of clone F49. Root tip squashes from F49 regenerants revealed the normal diploid chromosome number (2n=14). The frequency of regeneration within progeny from reciprocal crosses between F49 regenerants and several non-regenerative genotypes was 29%.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - BAP benzylaminopurine - KN kinetin - NAA -naphthaleneacetic acid     

Pollination in vitro: effects on the growth of pollen tubes,seed set and gametophytic self-incompatibility in Trifolium pratense L. and T. repens L.

Summary Growth of pollen tubes and seed set were compared after hand pollination in situ and in vitro in two self-incompatible species, Trifolium pratense and Trifolium repens. Adhesion of pollen grains to the stigma was greater in vitro for both species. After cross-pollination, in vitro culture gave a significant increase in the cumulative growth of pollen tubes in pistils of T. pratense compared to in situ conditions. After selfing in T. repens, pollen tube growth was significantly increased by in vitro culture of florets. Seed set after crossing in situ and in vitro was similar for both species. Seed set after selfing in vitro was not increased in T. pratense. Several genotypes of T. repens were classified as very good, good and poor selfers based on their capacity for seed set following selfing in situ. In vitro pollination increased self seed formation by 1.7-, 18.0- and 31.0-fold for each class, respectively. Ovules located nearest to the style were fertilized more often after selfing than after crossing.     

RNA-Seq analysis of compatible and incompatible styles of Pyrus species at the beginning of pollination

Plant Molecular Biology - At the early stage of pollination, the difference in gene expression between compatibility and incompatibility is highly significant about the pollen-specific expression...     

Growth and carbon partitioning in compatible and incompatible peach/plum grafts

The shoot growth of compatible ( Prunus persica L. Batsch cv. Springtime grafted on Prunus cerasifera L. Ehrh cv. Myrobolan P2032) and incompatible ( Prunus persica L. Batsch cv. Springtime grafted on Prunus cerasifera L. Ehrh cv. Myrobolan P18) peach/plum grafts was observed over a period of 100 days after grafting under controlled conditions. Leaf and root activities were determined by studying carbon assimilation and partitioning, leaf mineral contents and water relations. Shoot length and leaf number were not significantly affected in the incompatible combination during the first 55 days after grafting, but then, shoot growth rate was significantly reduced. Final total dry weights of the shoot were similar in both graft combinations. The incompatible combination did not show any water stress. Soluble sugar and starch contents increased in the leaves of the incompatible combination, accounting for about 36% of the increase of leaf dry weight per unit area. Photosynthesis was affected by the compatibility of the grafts. Leaf nitrogen content (% dry weight) fell in the incompatible graft combination 65 days after grafting. However, nitrogen content on an area basis was not affected. The possibility of nitrogen stress is discussed.     

Inheritance of somatic embryogenesis in red clover (Trifolium pratense L.)

 Red clover genotypes capable of regenerating plantlets in vitro from non-meristematic tissue-derived callus are rare. Selection for genotypes capable of somatic embryogenesis identified a clone comprised of a group of plantlets regenerated from a hypocotyl-derived callus culture on L2-based media and another group of plantlets originating from crown divisions of the epicotyl-derived plant. The callus-derived plants of this clone were highly regenerative when reintroduced to callus culture, but the epicotyl-derived plants produced nonregenerative callus cultures. F₁, F₂ and BC₁ populations were evaluated to determine the mode of inheritance of the regeneration trait. Reciprocal crosses did not differ, indicating a lack of maternal effects. Results were compatible with genetic control of regeneration by two complementary genes. We propose the genotype Rn1-Rn2- for regenerative plants. Three petiole segment explants were sufficient to evaluate regenerative ability in seedlings. Regenerative ability was often associated with abnormal leaf morphology in a few to several leaves. Received: 17 February 1998 / Accepted: 7 April 1998     

Flowering and gibberellins in a mutant red clover (Trifolium pratense L.)

Relationships between gibberellins and floral initiation were investigated in a conditional non-flowering mutant of red clover, Trifolium pratense. Untreated mutant plants will not flower under long-days, but will do so when certain GAs are applied. Gibberellins, A₃, A₁, A₇, and A₅ all resulted in both stem elongation and flowering whilst GA₄ produced the elongation only. Applications of GA₂₀, GA₈ and GA₁₃ under long-days had no detectable effect. Thus, by combining the use of the mutant with the application of different GAs, the correlation between the processes of stem elongation and floral initiation, which is normally strongly expressed in this species, was broken. Endogenous gibberellins shown to be present in normal plants were also found in the mutant genotype. Gibberellins alone were not sufficient to initiate floral development in the mutant, there being an essential element of interaction with long-days. These results are discussed in relation to the nature of the lesion in the mutant and the signal provided by the applied gibberellin.     

Pollen-Stigma Interaction in the Leguminosae: the Organization of the Stigma in Trifolium pratense L.

The intact stigma of Trifolium pratense possesses a smooth receptivesurface fringed by a few ranks of brush hairs. This surfaceis ensheathed by a thin (75–100 nm) but highly impermeablecuticle, which encloses four to five ranks of secretory cellsimmersed in their secretory products. Experimental single-grainpollinations show that pollen cannot become hydrated or germinateon the intact surface. The cuticle is ruptured when the floweris tripped; the secretion is released, and captured pollen-selfor cross-can then germinate. As in other papilionoid Leguminosae,this mechanism provides a guard against premature selling. Thesecretory cells are elongated; they remain in communicationthrough persistent pit-fields as the intercellular spaces fillwith secretion product. The secretion forms a lipid-rich emulsion,with a mucilaginous aqueous phase which reacts cytochemicallyfor protein and carbohydrate and has esterase activity. Duringthe early development of the stigma head, the cells possessa fine-structure appropriate to their secretory function, withabundant ribosomal and smooth endoplasmic reticulum, stratifiedor in the form of ramifying and anastomosing tubules, numerousmitochondria and a well developed Golgi system. Lipid globuli,partly invested in endoplasmic reticulum, are abundant in theyoung cells, but there is as yet no indication of how the lipidis transferred to the intercellular spaces during the secretoryperiod. As the stigma matures, the secretory cells become moribund. Leguminosae, Trifolium pratenseL., pollen-stigma interaction, self-incompatibility, autofertility, stigma secretory system, lipid secretion, cuticle permeability     

Changes in glucose and fructose level inNicotiana alata styles and ovaries accompanying compatible and incompatible pollen tube growth

Jestli?e byly pokusně květy den p?ed opylenim odděleny od rostliny a inkubovány ve vodě za teploty 25° C, projevil se v jejich pestících během následujících t?í dn? r?st pylových lá?ek v hladině glukosy a fruktosy. Prvý den po opylení bylo mno?ství těchto cukr? v apikálních ?ástech ?nělek z neopylených květ? vy??í, v basálních úsecích a v semeníeích naopak vět?inou ni??í ne? v p?íslu?ných ?ástech květ? opylených. V dal?ích dvou dnech do?lo i zde v p?ípadě opylení, a to p?edev?ím po allogamii, k silněj?ímu úbytku obou glycid?, tak?e po t?etím dnu bylo glukosy a fruktosy nejvíce v pestících neopylených, nejmáně po kompatibilním sprá?ení. Tento pokles byl nejvýrazněj?í v semeníeích, i kdy? do nich ani kompatibilní lá?ky je?tě nepronikly. V pokusech, kdy byly květy ponechány na rostlinách kultivovaných v polních podmínkách, nedo?lo v jejich pestících ani 80 hodin po kompatibilním opylení ke sní?ení obsahu glukosy a fruktosy. Z uvedených skute?ností lze vyvodit tyto záváry: R?st lá?ek ?nělkou vyvolá vá zvý?ený p?ísun glycid? do celých pestík?. Jak kompatibilní, tak inkompatibilní lá?ky vyu?ívají cukry z ?nělkováho pletiva. Oba tyto jevy jsou intenzívněj?í po allogamii ne? v p?ípadě inkompatibilní autogamie. Vzhledem k tomu, ?e kompatibilní lá?ky rostly normálně ?nělkou i za sní?ené hladiny glukosy a fruktosy, není absolutní zvý?ení obsahu těchto cukr? v pestících pro r? st lá?ek nezbytné. V opylených ?nělkách se mění poměr glukosa/fruktosa ve prospěch glukosy. Hodnota tohoto kvocientu se zvy?uje jak v apikálních, tak v basálních ?ásteeh ?ně1ek p?edev?ím po opylení kompatibilním. Tento jev je v souladu s hypothesou uvedenou d?ive (TUpý- 1959, 1960 podle ní? pylové lá?ky prodýchávají hlavnð sacharosu a z ní p?edev?ím její fruktofuranosovou slo?ku.     

白三叶和红三叶对人工融冻胁迫的生理响应差异

采用人工模拟融冻胁迫方法,通过测定白三叶(Trifolium repens)和红三叶(T.prat-ense)在融冻胁迫中叶片细胞膜透性、MDA含量、抗氧化酶(SOD、POD、CAT)活力、渗透调节物(脯氨酸、可溶性糖和蛋白质)含量变化,以揭示未来气候变化对三叶草的影响。结果表明,经历融冻胁迫循环后抗冻力强的白三叶植株能恢复生长,而抗冻力弱的红三叶枯萎死亡。在融冻阶段,两三叶草叶片细胞膜透性增大、抗氧化酶活力增高、MDA和渗透调节物含量大幅增加;在冻融阶段,两三叶草叶片细胞膜透性降低、MDA含量下降、抗氧化酶活力降低。但在融冻胁迫循环中,白三叶叶片POD和CAT活力高于红三叶,脯氨酸含量较红三叶高5倍,但细胞膜透性低于红三叶。白三叶在-5℃抗逆生理指标达到最大值,而红三叶在-10℃。白三叶对环境温度变化反应敏感,在-5℃通过快速激活抗氧化酶系统和积累渗透调节物以抑制膜脂过氧化和维护细胞水分平衡在融冻适应上起重要作用。白三叶具有较强的抗融冻能力,是未来值得应用推广的优良园林绿化植物。     

Integration of Linkage and Chromosome Maps of Red Clover ( Trifolium pratense L.)

Red clover (Trifolium pratense L.) is a forage legume and an allogamous diploid plant (2n = 14; 440 Mb). Here, we examine the 7 prometaphase chromosomes of red clover using fluorescence in situ hybridization (FISH) with ribosomal RNA sequences, pericentromeric and telomeric repeats, as well as bacterial artificial chromosome (BAC) clones. Position of hybridization signals and chromosome condensation patterns were quantified by the help of the chromosome image analysis system ver. 4.0 (CHIAS IV). Fourteen BAC clones belonging to linkage groups (LG) 1-7 hybridized to individual chromosomes 4, 2, 6, 5, 1, 7, and 3, respectively. Quantitative analysis of FISH mapping and chromosome analysis using CHIAS IV allowed us to construct a quantitative idiogram that constitutes the comprehensive chromosome map of red clover. Chromosomal positions of the 26S rDNA locus were detected at a heterozygous locus on chromosome 6 in the variety HR, and polymorphisms of rDNA loci were observed in other varieties, although chromosomal positions of some BAC clones did not vary among HR and other varieties. These results demonstrate chromosomal collinearity among allogamous red clover varieties. This integration of genetic linkage and quantitative chromosome maps should provide valuable insight into allogamous legume genetics.     

Pollen--stigma Interaction in the Leguminosae: the Secretory System of the Style in Trifolium pratense L.

The canal that traverses the upper part of the style of Trifoliumpratense is derived lysigenously. The core tissue of the veryyoung style consists of elongated cells similar to those ofthe transmitting tissue of solid-style families such as theSolanaceae; as the style matures, these cells separate to formthe canal, which receives secretions both from the core tissueand the inner wall cells. The early secretion of proteins intothe intercellular spaces is associated with the presence ofparamural bodies (lomasomes) in the adjacent cells. In the cellsin the immediate vicinity of the canal, vesicles, probably derivedfrom the Golgi system enlarge during later development and accumulatea protein-carbohydrate content, which is later passed into thecytoplasm where it forms densely packed fibrillar nodules. Withthe dissolution of the cell membranes, this material is passedinto the canal, where it is progressively diluted by continuedingress of water until the cavity reaches its final volume. Leguminosae, Trifolium pratense L., pollen—stigma interaction, self-incompatibility, stylar secretion, protein secretion