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Maize Gametophytic factor loci Ga3 through Ga11 modify reproductive barriers.

Gametophytic factor (Ga) barriers are maize (Zea mays ssp. mays) reproductive barriers controlled by molecular incompatibilities between pollen and silks. Twelve distinct Ga loci have been identified in maize populations since the first genetic evidence of a Ga barrier was reported in 1901. Of the twelve, however, only three have been validated by modern molecular, functional and genomic studies: Ga1, Ga2, and Tcb1. The remaining "higher" Ga loci, spanning Ga3 to Ga11, were reported in the historical literature, but their associated phenotypes segregated in unexpected ways or disappeared over subsequent generations. Here we introduce and explore the hypothesis that the higher Ga loci represent modifiers of Ga1, Ga2, and Tcb1 barrier functions. By revisiting the historical literature, we found that barrier phenotypes fall into two phenotypic and functional categories. Phenotypically, the two categories represented healthy pollen with a silk-length effect and unhealthy pollen without a silk-length effect. These phenotypic categories were supported by genomic data; we identified candidate genes in each higher Ga locus by comparing historical linkage mapping data to the corresponding genomic sequence of maize reference line B73. We discovered candidate genes related to two broad pathways: pollen tube growth and RNA-directed DNA methylation. We conclude that the past century of evidence aligns with our hypothesis that maize loci Ga3 through Ga11 modify rather than directly control Ga barriers. This brief investigation provides a starting point for geneticists and evolutionary biologists to explore how strong reproductive barriers are shaped by epistatic interactions.

Epistasis

[Spontaneous and induced sterility by ethyl methanesulfonate (EMS) in Nigella damascena L].

EMS-induced sterility could be very partially due to chromosomal aberrations appearing during male and female meiosis or even to mechanical abnormalities of the double fertilization. The sterility could be also related to diplontic origin (lethal factors or small deficiencies appearing in homozygous state on account of self-pollinization). Owing to our histological and genetical data, a female gametophytic origin could be mainly ascribed to EMS induced sterility. It could arise from a damage of the feeding function of the nucellus.

Chromosome Aberrations

The SPL-family transcription factor MpSPL3 orchestrates the proper regulation of vegetative and reproductive programs in Marchantia polymorpha.

SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) genes encode plant-specific transcription factors that are widely distributed across the plant kingdom. In angiosperms, the multimember SPL family regulates various biological processes, including vegetative-to-reproductive phase transition, inflorescence architecture, and lateral organ development. In contrast, the liverwort Marchantia polymorpha genome encodes only four SPL genes, with functional studies available only for microRNA-targeted members, MpSPL1 and MpSPL2. MpSPL1 was shown to control the meristem dormancy to modulate the thallus architecture, whereas MpSPL2 was found to promote the transition from vegetative-to-reproductive phase. Here, we investigate the impact of the MpSPL3 gene on M. polymorpha development. We demonstrate that MpSPL3 influences coordination of the vegetative growth and the reproductive phase transition. Knockout of MpSPL3 leads to strong growth retardation with disordered thallus morphology, reduced gemma cup number, and, most strikingly, complete loss of gametangiophore formation. Interestingly, overexpression of MpSPL3.2, the shorter isoform, has no detectable morphological effect, whereas the overexpression of MpSPL3.1, the longer isoform encoding a protein with an additional 61-aa long fragment, results in a delay in timing and reduced efficiency of gametangiophore production. Moreover, all the observed developmental abnormalities might be a consequence of the altered expression of genes essential for proper vegetative development and responsible for germ cell specification in MpSPL3 knockout and overexpression plants. Altogether, our findings demonstrate that MpSPL3 is important in regulating gametophyte development and ensuring reproductive success in M. polymorpha.

Marchantia

OsIDD6, an INDETERMINATE DOMAIN containing transcription factor in rice, plays an essential role in reproductive development.

INDETERMINATE DOMAIN containing proteins (IDD) are plant-specific transcriptional factors with a diverse range of roles in plants. Among the 15 IDD genes in rice, a staple food crop for the world, only about half have been functionally characterized. To elucidate the function of the remaining members, we created loss-of-function mutants using the CRISPR genome editing technique. Although no mutant exhibited obvious growth phenotypes, the Osidd6 mutant was completely sterile. By genetic crossing, we showed that both the male and female gametophytes were defective in the mutant. Histochemical staining and thin sectioning revealed that microspore development was compromised, likely due to a delay in tapetum degeneration. We also showed that meiosis was impaired in the mutant, resulting in defective megaspore development. Through a series of experiments, including transcriptome analysis, reverse transcription-quantitative polymerase chain reaction (RT-qPCR), in situ hybridization, β-glucuronidase (GUS) staining with promoter-GUS transgenic plants, yeast one-hybrid method, a dual-visible reporter assay, and transcriptional activity assay, we demonstrated that OsIDD6 is expressed in all cell types in the male and female reproductive organs and that the OsIDD6 protein directly regulates genes potentially having a role in meiosis and tapetum development. Since reproductive development is directly related to crop yield, OsIDD6 could be an important target for genetic manipulation in rice breeding.

Oryza

Genetic basis for broad interspecific compatibility in Solanum verrucosum.

Solanum verrucosum Schlechtendal (2x = 2n = 24) is unique among the clade 4 Solanum Sect Petota species. In addition to being one of the only fully self-compatible diploid potato species, S. verrucosum is the only clade 4 species that lacks prezygotic interspecific reproductive barriers. This allows S. verrucosum to accept pollen from a broad range of Solanum species and thereby serving as a genetic "bridge" between the cultivated or primary potato gene pool and distantly related wild relatives in the tertiary gene pool. The genetic mechanisms underlying self-compatibility in Solanum often underpin interspecific compatibility interactions, which in S. verrucosum, has been attributed to the lack of S-RNase expression. Using an interspecific F2 mapping population (n = 150), we investigated the genetic mechanisms responsible for the lack of interspecific reproductive barriers in S. verrucosum. This F2 population was evaluated for the ability to accept pollen from two clade 1, 1 EBN species (S. pinnatisectum and S. tarnii); from which two QTL for interspecific compatibility were identified on chromosomes 1 and 11, explaining 56.6% of the phenotypic variation observed. To identify the genetic basis of interspecific compatibility, we generated a chromosome-scale genome assembly of S. verrucosum MSII1813-2 and performed gene expression profiling of reproductive organs. Differential gene expression of S-RNase, located within the chromosome 1 QTL, confirmed the central role of the S-locus and specifically, S-RNase, in interspecific compatibility. Discovery of a non-S-locus QTL is consistent with previous findings that other non-S-locus factors are necessary for interspecific compatibility in S. verrucosum.

Solanum

[Effect of prolonged glucidic starvation on the contents of starch and tannins of Blechnum brasiliense L. gametophytes].

Some Blechnum brasiliense gametophytes have been maintained in darkness, in a purely mineral medium, for the purpose of investigating the effect of prolonged glucidic starving on the amount of tannins and starch that they contain. During the first two months the tannin contents undergoes a fast decrease. The starch variations are different : a transitory increase simultaneous with the fall in tannin; a decrease after the disappearance of 3/4 of the tannin contents; and lastly, a new amylogenese which precedes the prothallus necrosis. It seems that the gametophytes, in order to survive, degrade first their tannoidic reserves, and they utilize the starch that they contain only with difficulty. This behaviour can be explained by the distribution of the two metabolites; by the acid pH of the medium, which is not very propitious to the phosphorylases activity and lastly by the presence of chlorogenic acids, which are inhibitors of those enzymes.

Cells, Cultured

Spatial inheritance patterns across maize ears are associated with alleles that reduce pollen fitness.

Often, more pollen grains land on recipient flowers than there are ovules to fertilize. Consequently, the haploid male gametophyte engages in post-pollination competition, one way that pollen genotype can influence inheritance. The maize (Zea mays subsp. mays L.) inflorescence (ear), with its elongated stigma and style structures (silks), has a conspicuous spatial heterogeneity, with longer silks at the base of the ear than at the apex. To evaluate the hypothesis that alleles with reduced pollen fitness influence the spatial distribution of progeny genotypes along the ear, we developed an updated phenotyping platform that maps fluorescently marked mutant (Ds-GFP) kernel phenotypes on the ear via an implementation of the Faster R-CNN machine vision model (EarVision.v2) and a statistical pipeline that evaluates the relationship between kernel position and transmission ratio (EarScape). Our dataset (1384 ears) represents 58 Ds-GFP insertion alleles. None of the 48 alleles with Mendelian inheritance showed any significant spatial trend. In contrast, 50% of alleles with a pollen-specific transmission defect (5/10) exhibited significant spatial effects. An insertional mutant of the gene encoding a putative actin-binding protein, base-to-apex gradient1* (bag1*), is associated with decreased mutant transmission at the ear base relative to the apex. Surprisingly, a mutant allele of another pollen-expressed gene (Zm00001eb236740) generates the opposite trend, decreased mutant transmission toward the ear apex; and two mutant alleles of the sperm cell attachment factor gamete expressed2 (gex2) can produce ears with transmission highest at both base and apex. We conclude that pollen fitness mutants cause unexpectedly diverse spatial patterns of progeny genotypes.

Zea mays