Self-incompatibility and interspecific incompatibility: relationships in intra- and interspecific crosses of Zinnia elegans Jacq. and Z. angustifolia HBK (Compositae)

Summary Intraspecific and reciprocal interspecific crosses involving Zinnia angustifolia clones and Z. elegans lines showed that in both species, sporophytic self-incompatibility (SI) systems were present. Intensity of SI varied among clones and lines, and high self seed set was associated with a concomitant decrease in callose fluorescence in papillae and pollen tubes. Incomplete stigmatic inhibition of pollen germination and tube growth was observed in reciprocal interspecific crosses and associated with callose synthesis, suggesting S-gene activity. Seed set and progeny obtained following Z. angustifolia×Z. elegans matings was comparable to intraspecific compatible matings of Z. angustifolia although the rate of pollen tube growth through the style was slower. In Z. elegans × Z. angustifolia matings, additional prezygotic barriers were present and acted between pollen tube penetration of the stigma and syngamy. SI X SI interspecific incompatibility was essentially unilateral, with no embryos or progeny obtained when Z. elegans was the pistillate parent. It was hypothesized that nonfunctioning of Z. elegans × Z. angustifolia crosses was due to S-gene expression at the stigmatic surface and to other isolating mechanisms in the stylar or ovarian transmitting tissue.Scientific Article No. A-4448, Contribution No. 7439 of the Maryland Agricultural Experiment Station, Department of HorticultureA portion of this paper was presented in the report: Boyle TH, Stimart DP (1986) Incompatibility relationships in intra- and interspecific crosses of Z. elegans Jacq. and Z. angustifolia HBK (Compositae). In: Mulcahy D (ed) Biotechnology and ecology of pollen. Springer, New York     

Transitions between self-compatibility and self-incompatibility and the evolution of reproductive isolation in the large and diverse tropical genus Dendrobium (Orchidaceae)

Background and Aims The evolution of interspecific reproductive barriers is crucial to understanding species evolution. This study examines the contribution of transitions between self-compatibility (SC) and self-incompatibility (SI) and genetic divergence in the evolution of reproductive barriers in Dendrobium, one of the largest orchid genera. Specifically, it investigates the evolution of pre- and postzygotic isolation and the effects of transitions between compatibility states on interspecific reproductive isolation within the genus.Methods The role of SC and SI changes in reproductive compatibility among species was examined using fruit set and seed viability data available in the literature from 86 species and ∼2500 hand pollinations. The evolution of SC and SI in Dendrobium species was investigated within a phylogenetic framework using internal transcribed spacer sequences available in GenBank.Key Results Based on data from crossing experiments, estimations of genetic distance and the results of a literature survey, it was found that changes in SC and SI significantly influenced the compatibility between species in interspecific crosses. The number of fruits produced was significantly higher in crosses in which self-incompatible species acted as pollen donor for self-compatible species, following the SI × SC rule. Maximum likelihood and Bayesian tests did not reject transitions from SI to SC and from SC to SI across the Dendrobium phylogeny. In addition, postzygotic isolation (embryo mortality) was found to evolve gradually with genetic divergence, in agreement with previous results observed for other plant species, including orchids.Conclusions Transitions between SC and SI and the gradual accumulation of genetic incompatibilities affecting postzygotic isolation are important mechanisms preventing gene flow among Dendrobium species, and may constitute important evolutionary processes contributing to the high levels of species diversity in this tropical orchid group.     

Self-incompatibility inCrocus vernus subsp.vernus (Iridaceae)

Outcross, self- and mixed pollinations were performed inCrocus vernus subsp.vernus, a species with bicellular pollen, dry stigmas and hollow style. No differences were noted among the above pollinations concerning the germination of pollen and the growth of pollen tubes until the top of ovary. Within 45 min after pollinations 62% of pollen grains germinated. Pollen tubes penetrated the papilla cuticle extending along the papilla wall; on entry into stigmatic lobes they continued growth in the stylar secretion to ovarian locules. Here, however, self-pollen tubes failed to reach or to enter the ovule micropyle; while pollen tubes from either outcross- or mixed pollinations grew until fertilizing ovules. These observations gave evidence of a self-incompatibility system inCrocus, which appeared to be neutralized by mentor effect. The ovary as site of incompatibility response is discussed.     

A statistical analysis of some genetical,physiological and anatomical parameters of the development of in situ- and in vitro-grown ovules from intra- and interspecific crosses in the genus Beta

Summary Structural observations on in situ- and in vitro-grown ovules from different intra- and interspecific crosses in the genus Beta were tested using computer registration and statistical analysis. In intra- as well as interspecific crosses embryo development occurs with the same rate independent of growth conditions. Therefore, with respect to growth rate, embryo development seems to be highly autonomous, provided that nutrient influx into the ovule is sufficient. The endosperm develops significantly more rapidly during in vitro culture than in situ. A large degree of variability with respect to structural changes in the ovule tissue during in vitro culture is observed in and between the crosses. In general, the interspecific cross responds more rapidly on in vitro culture. In all crosses the embryo, although genetically identical to the suspensor, shows a higher degree of response to in vitro culture than the embryo itself. Early suspensor degeneration in the interspecific cross is the only observed difference between the crosses, which could explains the lack of root formation in seedlings of interspecific hybrids. The use of statistical analysis on the anatomical parameters used to compare treatment as well as crosses has proven to be an efficient and novel approach to plant reproduction biology.     

Unilateral incompatibility within the brassicaceae: further evidence for the involvement of the self-incompatibility (S)-locus

Unilateral pollen-pistil incompatibility within the Brassicaceae has been re-examined in a series of interspecific and intergeneric crosses using 13 self-compatible (SC, Sc) species and 12 self-incompatible (SI) species from ten tribes. SC x SC crosses were usually compatible, SI x SC crosses showed unilateral incompatibility, while SI x SI crosses were often incompatible or unilaterally incompatible. Unilateral incompatibility (UI) is shown to be overcome by bud pollination or treating stigmas with cycloheximide — features in common with self-incompatibility. Treating stigmas with pronase prevents pollen tubes from penetrating the stigma in normally compatible intra-and interspecific pollinations. The results presented show that the presence of an incompatibility system is important in predicting the outcome of interspecific and intergeneric crosses and, combined with the physiological similarities between UI and SI, would suggest an involvement of the S-locus in UI.     

Relationships of pollen size,pistil length and pollen tube growth rates in Rhododendron and their influence on hybridization

Summary Pollen size and pistil length data have been collected for 93 species of Rhododendron (Ericaceae) belonging to a number of different subgeneric taxa. For a sample of eight species in section Vireya, pollen tube growth in the style after selfor interspecific pollination has been quantified. Pollen volume and the time taken for pollen tubes to reach the ovary were both related to pistil length. Pollen-tube growth rates were generally greater for species with longer pistils and larger pollen. Increasing temperature increased the rate of pollen-tube growth. There was no detectable effect of pollen tube density on tube growth rate in the style. After interspecific pollinations tube growth rates in foreign styles could be faster or slower than in self styles. A semisterile individual with two viable pollen grains per tetrad and a plant grafted as scion to a longer-styled stock both showed more rapid pollen-tube growth than expected on the basis of pistil size. Data collected for 26 species in section Vireya showed that where extreme disparity of pollen/pistil size causes failure of interspecific crosses, one or more bridging species with intermediate pollen/pistil size can generally be selected.     

An interpretation of genomic balance in seed formation in interspecific hybrids of Solanum

Summary To investigate the mechanisms of seed failure in intraspecific and interspecific crosses of Solanum two diploid, S. commersonii and Group Phureja, and one tetraploid species, S. acaule, species were crossed and the seeds were analyzed for embryo and endosperm development. Many seeds of certain crosses observed seven days after pollinations were found to contain abnormal embryos and degenerating endosperms. In some cases seeds contained an embryo with no endosperm, or an endosperm with no embryo. Other interspecific crosses which were predicted to fail actually produced seeds with normally developed embryos and endosperms. To further characterize the intra- and interspecific embryos and endosperms the nuclear DNA was measured. There are several ways to explain the ploidy levels of embryos and endosperms among the crosses, the occurrence of unreduced gametes in some cases and genomic instability in other cases. The latter resulted in chromosome loss at meiotic and mitotic divisions. Genomic balance in interspecific seeds is critical to both embryo and endosperm development.Scientific Journal Series Article No. 14636 of the Minnesota Experiment Station     

Pollen-pistil interactions in Populus: β-galactosidase activity associated with pollen-tube growth in the crosses P. nigra x P. nigra and P. nigra x P. alba

Summary In order to understand the nature of interspecific barriers in Populus, we have explored pollen/pistil interactions in intra- and interspecific crosses Populus nigra x P. nigra and P. nigra x P. alba. The kinetics of pollen-tube growth demonstrated that P. nigra and P. alba pollen tubes have distinct behavioral patterns inside P. nigra pistils. P. alba pollen tubes exhibit an unique S-shaped growth curve and an arrested growth site near the sylodium. P. nigra pollen tubes exhibit two growth phases, in the stigmatic tissues and in the ovarian cavity respectively. P. nigra and P. alba curves diverge 5 h after controlled pollination and could be related to a change in the physiology of the P. nigra pollen tube, which shifts from an autotrophic to a heterotrophic type of nutrition. Protein analysis of pollinated stigmatic extracts (0,6 and 20 h after pollination) revealed qualitative and quantitative differences that are related to the presence of either P. nigra or P. alba pollen tubes inside the stigmatic tissues. Increasing numbers of protein bands were detectable from 0 to 20 h after pollination only in intraspecific cross. Glycoproteins were detected, and the differences observed were dependent of the cross. -Galactosidase activity was found in pollinated stigmas, but an increase in its activity (one isozyme of pHi 4.2) between 6 h and 20 h after pollination was detected only in the intraspecific cross. This enzyme could play a role in heterotrophic pollen-tube nitrition, and its activity could be the final result of a series of interactions started by the initial pollen-stigma dialog.     

Self-incompatibility of Zinnia angustifolia HBK (compositae)

Summary Visible light and UV epifluorescence microscopy were used to assess self-incompatibility (SI) in Zinnia angustifolia clones. Pistils were fixed 24 h after pollination and stained either with aniline blue in lactophenol (visible light microscopy) or decolorized aniline blue (fluorescence microscopy). Percentage of florets with embryos 21 days following pollination (% embryo set) was used as a control. Embryo set following self- or incompatible cross-pollinations ranged from 0% to 9.9%, whereas compatible crosses yielded 55.5%–87.1% embryo set. Observations using visible light microscopy indicated pollen load and number of germinated grains were significantly higher for compatible compared to incompatible crosses, and both variables were positively correlated (r = 0.89–0.96) to % embryo set. Examinations with UV epifluorescence microscopy revealed pollen load was higher and little or no callose accumulated in stigmatic papillae following compatible crosses, whereas for incompatible crosses, pollen load was low and callose lenticules were deposited in stigmatic papillae; the correlation between pollen load and % embryo set was 0.89. The intensity of callose fluorescence of the pollen tube-papillae attachment sites was quantitatively measured via micro spectrofluorometry. Callose fluorescence intensity ranged from 47.9% to 62.6% for incompatible and from 6.4% to 9.9% for compatible crosses, and was negatively correlated (r= — 0.95) with % embryo set. Microscopal techniques permit rapid assessment of SI and may be used routinely when each observed or measured parameter is highly correlated to the incompatibility response.Publication no. 2898 of the Massachusetts Agricultural Experiment Station.     

Interspecific incompatibility due to stylar barriers in tuber-bearing and closely related non-tuber-bearing Solanums

Summary The phenomenon of interspecific incompatibility between various wild tuber-bearing and closely related non-tuber-bearing Solanum species was studied. One area of investigation included an examination of possible protein interactions in the incompatibility reaction using SDS electrophoresis. Pollen tube inhibition and morphology were examined in conjunction with biochemical analysis. Two sets of crosses were examined: interspecific tuber-bearing species crosses and interspecific tuber-bearing × non-tuber-bearing species crosses. These crosses had consistent pollen tube inhibition in the upper one-third of the style. The upper third of the styles of incompatibly pollinated, compatibly pollinated, and unpollinated styles was studied under fluorescence microscopy to observe pollen tube growth and morphology. Interspecific tuber-bearing × non-tuber-bearing species crosses demonstrated consistent pollen tube inhibition just below the stigma with frequent pollen tube swelling and bursting and extensive callose deposition along the pollen tube wall. Interspecific tuber-bearing species crosses had pollen tube inhibition further down the style with pollen tube tip tapering and extensive callose deposition. Stylar proteins of the lower two-thirds of the styles were analyzed with SDS electrophoresis. No unique protein differences were found to be specifically associated with the interspecific incompatibility reaction in this portion of the style.Cooperative investigation of the U.S. Department of Agriculture, Agricultural Research Service, and the Wisconsin Experiment Station. Supported in part by the USDA, Cooperative States Research Service Competitive grant no. 83-CRCR-1-1253     

Unilateral incongruity in crosses involvingLycopersicon pennellii andL. esculentum is distinct from self-incompatibility in expression,timing and location

Both interspecific and intraspecific mechanisms restrict the exchange of genes between plants. Much research has focused on self incompatibility (SI), an intraspecific barrier, but research on interspecific barriers lags behind. We are using crosses betweenLycopersicon esculentum andL. pennellii as a model with which to study interspecific crossing barriers. The crossL. esculentum×L. pennellii is successful, but the reciprocal cross fails. Since the cross can be successfully made in one direction but not the other, gross genomic imbalance or chromosomal abnormality are precluded as causes. We showed that the lack of seed set observed in the crossL. pennellii×L. esculentum is due to the inability of pollen tubes to grow more than 2–3 mm into the style, whereas S1 crosses show continued slow pollen tube growth but, also, fail to set seed. These results indicate that the unilateral response is a barrier distinct from SI, differing from SI in the timing and location of expression in the style. We therefore suggest that this unilateral response in theL. pennellii×L. esculentum cross is more accurately referred to as unilateral incongruity (UI) rather than interspecific incompatibility. Periclinal chimeras were used to determine the tissues involved in UI. The results of crosses with the available chimeras indicate that the female parent must beL. pennellii at either LI (layer 1) or both LI and LII (layer 2) and the male parent must beL. esculentum at either LII or both LI and LII to observe UI similar to that seen in theL. pennellii×L. esculentum cross. Pollinations with a mixture of pollen fromL. pennellii and from transgenicL. esculentum plants harboring a pollen-specific GUS reporter gene marker were used to ascertain whether the growth of the pollen tubes of either species was modified as a possible means of overcoming UI. We found no evidence of communication between the two types of pollen tubes to either enhance or restrict all pollen tube growth.     

S RNase and Interspecific Pollen Rejection in the Genus Nicotiana: Multiple Pollen-Rejection Pathways Contribute to Unilateral Incompatibility between Self-Incompatible and Self-Compatible Species

In self-incompatible (SI) plants, the S locus acts to prevent growth of self-pollen and thus promotes outcrossing within the species. Interspecific crosses between SI and self-compatible (SC) species often show unilateral incompatibility that follows the SI x SC rule: SI species reject pollen from SC species, but the reciprocal crosses are usually compatible. The general validity of the SI x SC rule suggests a link between SI and interspecific pollen rejection; however, this link has been questioned because of a number of exceptions to the rule. To clarify the role of the S locus in interspecific pollen rejection, we transformed several Nicotiana species and hybrids with genes encoding SA2 or SC10 RNase from SI N. alata. Compatibility phenotypes in the transgenic plants were tested using pollen from three SC species showing unilateral incompatibility with N. alata. S RNase was implicated in rejecting pollen from all three species. Rejection of N. plumbaginifolia pollen was similar to S allele-specific pollen rejection, showing a requirement for both S RNase and other genetic factors from N. alata. In contrast, S RNase-dependent rejection of N. glutinosa and N. tabacum pollen proceeded without these additional factors. N. alata also rejects pollen from the latter two species through an S RNase-independent mechanism. Our results implicate the S locus in all three systems, but it is clear that multiple mechanisms contribute to interspecific pollen rejection.     

Interspecific crosses in the genus Tulipa L.: identification of pre-fertilization barriers

 Pollen tube growth in the pistil and pollen tube penetration of ovules have both been studied in crosses between cultivars from Tulipa gesneriana L. and 12 tulip species from all eight sections of the genus Tulipa to identify pre-fertilization barriers. Depending on the cross, pollen tubes grew as far as the stigma or the style or continued growing down into the ovary. Pollen tubes penetrated none or only a few percent of the ovules of some crosses, despite the presence of many pollen tubes in the ovary. In other crosses, from which no or only a few hybrids have been obtained after seed maturation on the plant, pollen tube penetration was found in up to 79% of the ovules. Apparently, various kinds of barriers preventing fertilization or normal embryogenesis occur in interspecific tulip crosses. Received: 26 July 1996 / Revision accepted: 31 January 1997     

Inheritance of some Mendelian factors in intra- and interspecific crosses between Setaria italica and Setaria viridis

Summary The inheritance of seed coat color, pericarp color, polyphenoloxidase activity and bristle, glume, collar, and leaf-base anthocyanic colorations was investigated using intra- and interspecific crosses between Setaria italica and S. viridis. The results were compared to inheritance results obtained by previous authors. In most cases, the inheritance is simple (one or two loci) and data from different crosses (intra- and interspecific) and from different authors can be compared. Two sets of two characters were found to share common loci: the polyphenoloxidase locus is one of the loci responsible for seed coat color, and bristle and glume color are determined by the same two loci. The evolutionary significance of these results is discussed.     

Reproductive systems and crossing potential in three species ofBulbophyllum (Orchidaceae) occurring in Brazilian ‘campo rupestre’ vegetation

Experimental self-, cross- and interspecific pollinations were carried out inBulbophyllum weddellii, B. involutum andB. ipanemense (Orchidaceae). The last two species are closely related, butB. weddellii is somewhat more distant. The three species are self-compatible, with pollination by a vector necessary for fruit formation. In all crossing types, high rates of fruit abortion and formation of seeds without embryos were observed, particularly in interspecific crosses that involvedB. weddellii. Crosses between the more closely relatedB. involutum andB. ipanemense produced fruit formation rates and seed viability similar to those obtained in intraspecific crosses. These results agree with current hypotheses that suggest that interspecific crossing rates reflect phylogenetic proximity. Examination of pollen tube growth and aborted fruits suggests that a series of factors may be involved in the high abortion rate for fruits and large proportion of seeds without embryos. The low interspecific fertility noted forB. weddellii andB. involutum is important in the maintenance of isolation between these species that are sympatric, flower in the same period and share the same pollinators.     

Pollen tube expression of pseudo-self-compatibility (PSC) inPetunia hybrida

Summary AnS _1.1 self-incompatible (SI) petunia plant which showed atypical seed set was found in an I₇ population. This plant showed a strong SI reaction when selfed but produced varying amounts of seed when used as the seed parent in crosses with unrelated individuals homozygous for the sameS allele. Reciprocal crosses yielded no seed indicating that the reaction was a stylar response. Self seed obtained by high temperature treatments produced 18 plants, all of which exhibited the parental characteristics, the ability to reject self pollen but accept, to varying degrees, pollen bearing the sameS allele from unrelated plants. Several petunias homozygous forS ₁, and exhibiting various levels of PSC as determined by self seed set, progeny tests and temperature treatments, were used as pollen parents. The mean seed set of these crosses produced a ranking of the pollen parents which reflected the PSC levels obtained by other methods. The behavior of the F₁ and F₂ populations suggests that the pollen discriminating ability may be a simply inherited, dominant character in these plants. The styles of these unusual petunias illustrate the participation of the pollen tube in determining PSC.Scientific Journal Series Paper Number 10.479 of the Minnesota Agricultural Experiment Station     

Post-pollination hybridization barriers in <Emphasis Type="Italic">Nicotiana</Emphasis> section <Emphasis Type="Italic">Alatae</Emphasis>

Nicotiana section Alatae contains eight species with variable flower sizes and morphologies. Section members readily hybridize in the glasshouse, but no hybrids have been observed in natural sympatric and parapatric populations. To investigate interspecific crossing relationships with respect to mechanisms preventing hybridization, all members of section Alatae were intercrossed in a complete diallel. We found positive correlation between the pistil length of the pollen donor and interspecific seed set relative to the conspecific cross. Pollen tube growth rate and pollen donor pistil length were positively correlated as well. Furthermore, pollen from short-pistil members of section Alatae could only grow a maximum distance proportional to, but greater than, their own pistil lengths. Our results show that pollen tube growth capacity (i.e., rate and distance), provides a hybridization barrier in long-pistil species × short-pistil species crosses. We also found another hybridization barrier not specifically related to pollen tube growth capacity in short-pistil species × long-pistil species. Taken together, these barriers can generally be described by a ‘pistil-length mismatch’ rule; in section Alatae, pollen has the most success fertilizing ovules from species with pistil lengths similar to their own. This rule could contribute to hybridization barriers in Section Alatae because the species display dramatically different pistil lengths. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Geographical variation in postzygotic isolation and its genetic basis within and between two Mimulus species

The aim of this study is to investigate the evolution of intrinsic postzygotic isolation within and between populations of Mimulus guttatus and Mimulus nasutus. We made 17 intraspecific and interspecific crosses, across a wide geographical scale. We examined the seed germination success and pollen fertility of reciprocal F₁ and F₂ hybrids and their pure-species parents, and used biometrical genetic tests to distinguish among alternative models of inheritance. Hybrid seed inviability was sporadic in both interspecific and intraspecific crosses. For several crosses, Dobzhansky–Muller incompatibilities involving nuclear genes were implicated, while two interspecific crosses revealed evidence of cytonuclear interactions. Reduced hybrid pollen fertility was found to be greatly influenced by Dobzhansky–Muller incompatibilities in five out of six intraspecific crosses and nine out of 11 interspecific crosses. Cytonuclear incompatibilities reduced hybrid fitness in only one intraspecific and one interspecific cross. This study suggests that intrinsic postzygotic isolation is common in hybrids between these Mimulus species, yet the particular hybrid incompatibilities responsible for effecting this isolation differ among the populations tested. Hence, we conclude that they evolve and spread only at the local scale.     

Intergeneric hybridization between Diplotaxis siifolia,a wild species and crop brassicas

Summary Attempts were made to obtain intergeneric hybrids between Diplotaxis siifolia, a wild species, and cultivars of Brassica (B. campestris, B. juncea, and B. napus). The crosses showed unilateral incompatibility. When the wild species was used as female parent, pollen germination and pollen tube growth were normal, but hybrid seeds aborted due to post-fertilization barriers. Reciprocal crosses (cultivars as female parent) showed strong pre-fertilization barriers; although pollen grains showed germination, pollen tubes failed to enter the stigma. Hybrids were realized in two of the crosses, D. siifolia x B. juncea and D. siifolia x B. napus, through ovary culture. The hybrids were multiplied in vitro by multiplication of axillary shoots, or somatic embryogenesis. Detailed studies were carried out on the hybrid D. siifolia x B. juncea. F₁ hybrids had shrivelled anthers and were pollen sterile. Amphiploids of this hybrid showed 60% pollen fertility and produced seeds upon self-pollination as well as backcross pollination with the pollen of B. juncea.     

小报春与岩生报春种间杂交亲和性研究

以报春花属报春花组的小报春(Primula forbesii)和指叶报春组的岩生报春(Primula saxatilis)为亲本,对种间杂交的结实性及花粉管行为进行了观察。结果表明:小报春与岩生报春种间杂交表现为不亲和,正反交组合正常结实率为0,但正交、反交组合花粉在柱头表面的萌发和花粉管伸长过程有明显差异,小报春花粉授粉4h后可以在岩生报春柱头表面萌发,但花粉管伸长的速度明显比对照组[岩生报春(P)×岩生报春(T)、岩生报春(T)×岩生报春(P)]慢,并且花粉管生长弯曲,授粉192h后花粉管仍未到达子房;岩生报春的花粉可以在小报春柱头上正常萌发,授粉48h后花粉管到达子房;4个对照授粉组合均分别于24h(短花柱为母本)、48h(长花柱为母本)时完成受精过程。研究表明,岩生报春×小报春杂交存在受精前障碍,小报春×岩生报春杂交亲和性较好,并可通过幼胚拯救的方法获得组间杂种后代;花柱长度可能是影响种间杂交结实能力的因素之一。