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A plant can be thought of as a colony comprising numerous growth buds, each developing to its own rhythm. Such lack of synchrony impedes efforts to describe core principles of plant morphogenesis, dissect the underlying mechanisms, and identify regulators. Here, we use the tiniest known angiosperm to overcome this challenge and provide an ideal model system for plant morphogenesis. We present a detailed morphological description of the monocot Wolffia australiana, as well as high-quality genome information. Further, we developed the Plant-on-Chip culture system and demonstrate the application of advanced technologies such as snRNA-seq, protein structure prediction, and gene editing. We provide proof-of-concept examples that illustrate how W. australiana can open a new horizon for deciphering the core regulatory mechanisms of plant morphogenesis.SignificanceWhat is the core morphogenetic process in angiosperms, a plant like a tree indeterminately growing, or a bud sequentially generating limited types of organs? Wolffia australiana, one of the smallest angiosperms in the world may help to make a distinction. Wolffia plantlet constitutes of only three organs that are indispensable to complete life cycle: one leaf, one stamen and one gynoecium. Before the growth tip is induced to flower, it keeps branching from the leaf axil and the branches separate from the main plantlet. Here we present a high-quality genome of W. australiana, detailed morphological description, a Plant-on-Chip cultural system, and some principle-proof experiments, demonstrating that W. australiana is a promising model system for deciphering core developmental program in angiosperms.

Chromosome segregation must be under strict regulation to maintain chromosome euploidy and stability. Cell Division Cycle 20 (CDC20) is an essential cell cycle regulator that promotes the metaphase-to-anaphase transition and functions in the spindle assembly checkpoint, a surveillance pathway that ensures the fidelity of chromosome segregation. Plant CDC20 genes are present in multiple copies, and whether CDC20s have the same functions in plants as in yeast and animals is unclear, given the potential for divergence or redundancy among the multiple copies. Here, we studied all three CDC20 genes in rice (Oryza sativa) and constructed two triple mutants by clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9-mediated genome editing to explore their roles in development. Knocking out all three CDC20 genes led to total sterility but did not affect vegetative development. Loss of the three CDC20 proteins did not alter mitotic division but severely disrupted meiosis as a result of asynchronous and unequal chromosome segregation, chromosome lagging, and premature separation of chromatids. Immunofluorescence of tubulin revealed malformed meiotic spindles in microsporocytes of the triple mutants. Furthermore, cytokinesis of meiosis I was absent or abnormal, and cytokinesis II was completely prevented in all mutant microsporocytes; thus, no tetrads or pollen formed in either cdc20 triple mutant. Finally, the subcellular structures and functions of the tapetum were disturbed by the lack of CDC20 proteins. These findings demonstrate that the three rice CDC20s play redundant roles but are indispensable for faithful meiotic chromosome segregation and cytokinesis, which are required for the production of fertile microspores.

Meiotic recombination may occur more frequently in some regions of the eukaryotic genome. These regions are referred to as meiotic recombination hotspot loci. Genetic studies in the yeast have revealed that these hotspots contain special sites for recombination initiation and result in heterogeneous distribution of recombination (along the chromosome length or across the genome). However, the data for recombination hotspot is so far limited to organisms such as fungi, maize and human. In the present paper, we listed several typical approaches to characterizing recombination hotspots in different organisms, summarizing current progress in mapping of the meiotic recombination hotspots and discussing the factors and mechanisms for activation of eukaryotic meiotic recombination. Some unresolved questions and future prospects were also discussed.

Rice calli derived from anther culture were used as recipient to transfer a rice blight resistance gene, Xa21, into a japonica rice variety, Taipei 309, via Agrobacterium-mediated transformation. Seven green transgenic plants, including one mixoploid, two haploid, and four diploid plants, were regenerated. PCR, Southern blot, FISH and blight resistance analysis all indicated that Xa21 gene has been integrated into the T0 plant genomes. T1 generations of the four diploid T0 plants were further investigated for resistance segregation. Chi2 test showed that two T1 populations segregated with a ratio of 3:1, indicating that a single copy of Xa21 gene was integrated into the genome, whereas the segregation ratios of the other two T1 populations were non-Mendelian. Therefore, the four diploid transgenic plants should be heterozygous diploids.

Rice (Oryza sativa L.) is one of the most important cereal crops in the world. The chromosomes of rice are relatively small in size. With the help of fluorescence in situ hybridization (FISH), several rice DNAs have been localized on rice chromosomes. 45S rDNA and 5S rDNA are encoding sequences for ribosomal RNA synthesis. For detecting the chromosomes related to 45S rDNA and 5S rDNA, the digoxigenin-dUTP labeled probe DNA was probed to the prometaphase chromosomes which were prepared from the root tips harvested from an indica rice variety, Zhongxian 3037. For identification of the chromosomes, slides were stained with Giemsa before FISH. With the improved FISH protocol 45S rDNA was clearly detected on two pairs of chromosomes which are usually found to be attached to nucleolus. The signal size between two pairs of the chromosomes was different. According to the characteristics of the chromosomes, the chromosomes with bigger signal were chromosome 9 and the other pair were chromosome 10. They were also discriminated from arm ratio. The signals of 5S rDNA were relatively small but clear. According to the size and arm ratio of the chromosome with FISH signals, the 5S rDNA was mapped on the short arm of chromosome 11, very close to the centromere region.

The aneuploid with isochromosome or telochromosome is ideal material for exploring the position of centromere in lingkage map. For obtaining these aneuploids in rice, the primary trisomics from triplo-1 to triplo-12 and the aneuploids derived from a triploid of indica rice variety Zhongxian 3037 were carefully investigated. From the offsprings of triplo-10, a primary trisomic of chromosome 10 of the variety, an isotetrasomic "triplo-10-1" was obtained. Cytological investigation revealed that a pair of extra isochromosomes of triplo-10-1 were come from the short arm of chromosome 10. In the offsprings of the isotetrasomic, a secondary trisomic "triplo-10-2", in which the extra- chromosome was an isochromosome derived from the short arm of chromosome 10, was identified. With the isotetrasomic, secondary trisomic, primary trisomic and diploid of variety Zhongxian 3037, different molecular markers were used for exploring the position of the centromere of chromosome 10. Based on the DNA dosage effect, it was verified that the molecular markers G1125, G333 and L169 were located on the short arm, G1084 and other 16 available molecular markers were on the long arm of chromosome 10. So the centromere of chromosome 10 was located somewhere between G1125 and G1084 according to the RFLP linkage map given by Kurata et al[1]. The distance from G1125 to G1084 was about 3.2 cM.

Segregation distortion is a quite common phenomenon in living species and thought to be a potent evolutional force. The main reasons of distorted segregation ratios are responsible for the selection of gametes or sporophytes. In this study, two extreme segregation distortions from the progenies of lmi x 02428 and d6 x 93-11 were identified. The segregation ratio of molecular markers tightly linked with LMI and D6 genes were analyzed and skew segregation were found in the markers tested which were indicated by significant deviation from the expected Mendelian segregation ratio(1:2:1). The segregation distorted regions were detected between molecular markers ST8 and ST8-2 near the centromere of chromosome 8, and ST7-1 and ST7-3 near telomere of chromosome 7, respectively. Meanwhile, the results indicated that segregation distortion had related with the different crossed combinations.