Overexpression Populus d-Type Cyclin Gene PsnCYCD1;1 Influences Cell Division and Produces Curved Leaf in Arabidopsis thaliana

d-type cyclins (CYCDs) are a special class of cyclins and play extremely important roles in plant growth and development. In the plant kingdom, most of the existing studies on CYCDs have been done on herbaceous plants, with few on perennial woody plants. Here, we identified a Populus d-type cyclin gene, PsnCYCD1;1, which is mainly transcribed in leaf buds and stems. The promoter of PsnCYCD1;1 activated GUS gene expression and transgenic Arabidopsis lines were strongly GUS stained in whole seedlings and mature anthers. Moreover, subcellular localization analysis showed the fluorescence signal of PsnCYCD1;1-GFP fusion protein is present in the nucleus. Furthermore, overexpression of the PsnCYCD1;1 gene in Arabidopsis can promote cell division and lead to small cell generation and cytokinin response, resulting in curved leaves and twisted inflorescence stems. Moreover, the transcriptional levels of endogenous genes, such as ASs, KNATs, EXP10, and PHB, were upregulated by PsnCYCD1;1. Together, our results indicated that PsnCYCD1;1 participates in cell division by cytokinin response, providing new information on controlling plant architecture in woody plants.

Complex haploinsufficient interaction between APC11 and CYCLIN A1;2/TARDY ASYNCHRONOUS MEIOSIS in embryo development and seed germination in Arabidopsis

Background: Complex haploinsufficiency is characterized by individuals with two heterozygous loci producing a phenotype that is not seen in either of the corresponding single-locus heterozygous individuals. The mutants of the anaphase-promoting complex/cyclosome (APC/C) subunit gene APC11 and the mitotic cyclin gene CYCLIN A1;2/TARDY ASYNCHRONOUS MEIOSIS (TAM) in Arabidopsis thaliana are embryo-lethal and defective in meiosis, respectively, but their heterozygous single mutants do not exhibit defective embryo development and seed germination. Methods: Crosses between two heterozygous apc11 mutant alleles and two homozygous tam mutant alleles, and between two heterozygous apc11 mutant alleles and a TAM:TAM-GFP line were conducted. Phenotypes of the F1 seeds were analyzed by light microscopy. Results: We found that F1 embryos from the crosses between heterozygous apc11-1 (APC11/apc11-1) and homozygous tam-2 (tam-2/tam-2) or between APC11/apc11-2 and tam-2/tam-2 were morphologically normal but all the seeds failed to germinate. F1 embryos from the crosses between APC11/apc11-2 and tam-1/tam-1 (weaker allele than tam-2) produced morphologically normal seeds that germinated to form mature plants. However, F1 embryos from the crosses between APC11/apc11-1 and tam-1/tam-1 were abnormal and the seeds failed to germinate. Moreover, F1 embryos from the crosses between APC11/apc11-1 and a TAM:TAM-GFP line were arrested at early developmental stages while F1 embryos from the crosses between APC11/apc11-2 and the TAM:TAM-GFP line appeared fully developed but the seeds failed to germinate. Conclusions: Our observations indicate that the apc11 and tam mutants have an allele-dependent complex haploinsufficient relationship in embryo development and seed germination.

Genome-Wide Analysis of the Cyclin Gene Family and Their Expression Profile in Medicago truncatula

Cyclins, together with highly conserved cyclin-dependent kinases (CDKs), play an important role in the process of cell cycle in plants, but less is known about the functions of cyclins in legume plants, especially Medicago truncatula. Our genome-wide analysis identified 58, 103, and 51 cyclin members in the M. truncatula, Glycine max, and Phaseolus vulgaris genomes. Phylogenetic analysis suggested that these cyclins could be classified into 10 types, and the CycB-like types (CycBL1-BL8) were the specific subgroups in M. truncatula, which was one reason for the expansion of the B-type in M. truncatula. All putative cyclin genes were mapped onto their own chromosomes of each genome, and 9 segmental duplication gene pairs involving 20 genes were identified in M. truncatula cyclins. Determined by quantitative real-time PCR, the expression profiling suggested that 57 cyclins in M. truncatula were differentially expressed in 9 different tissues, while a few genes were expressed in some specific tissues. Using the publicly available RNAseq data, the expression of Mtcyclins in the wild-type strain A17 and three nodule mutants during rhizobial infection showed that 23 cyclins were highly upregulated in the nodulation (Nod) factor-hypersensitive mutant sickle (skl) mutant after 12 h of rhizobium inoculation. Among these cyclins, six cyclin genes were also specifically expressed in roots and nodules, which might play specific roles in the various phases of Nod factor-mediated cell cycle activation and nodule development. Our results provide information about the cyclin gene family in legume plants, serving as a guide for further functional research on plant cyclins.

Phylogenetic analysis of cell-cycle regulatory proteins within the Symbiodiniaceae

In oligotrophic waters, cnidarian hosts rely on symbiosis with their photosynthetic dinoflagellate partners (family Symbiodiniaceae) to obtain the nutrients they need to grow, reproduce and survive. For this symbiosis to persist, the host must regulate the growth and proliferation of its symbionts. One of the proposed regulatory mechanisms is arrest of the symbiont cell cycle in the G1 phase, though the cellular mechanisms involved remain unknown. Cell-cycle progression in eukaryotes is controlled by the conserved family of cyclin-dependent kinases (CDKs) and their partner cyclins. We identified CDKs and cyclins in different Symbiodiniaceae species and examined their relationship to homologs in other eukaryotes. Cyclin proteins related to eumetazoan cell-cycle-related cyclins A, B, D, G/I and Y, and transcriptional cyclin L, were identified in the Symbiodiniaceae, alongside several alveolate-specific cyclin A/B proteins, and proteins related to protist P/U-type cyclins and apicomplexan cyclins. The largest expansion of Symbiodiniaceae cyclins was in the P/U-type cyclin groups. Proteins related to eumetazoan cell-cycle-related CDKs (CDK1) were identified as well as transcription-related CDKs. The largest expansion of CDK groups was, however, in alveolate-specific groups which comprised 11 distinct CDK groups (CDKA-J) with CDKB being the most widely distributed CDK protein. As a result of its phylogenetic position, conservation across Symbiodiniaceae species, and the presence of the canonical CDK motif, CDKB emerged as a likely candidate for a Saccharomyces cerevisiae Cdc28/Pho85-like homolog in Symbiodiniaceae. Similar to cyclins, two CDK-groups found in Symbiodiniaceae species were solely associated with apicomplexan taxa. A comparison of Breviolum minutum CDK and cyclin gene expression between free-living and symbiotic states showed that several alveolate-specific CDKs and two P/U-type cyclins exhibited altered expression in hospite, suggesting that symbiosis influences the cell cycle of symbionts on a molecular level. These results highlight the divergence of Symbiodiniaceae cell-cycle proteins across species. These results have important implications for host control of the symbiont cell cycle in novel cnidarian–dinoflagellate symbioses.

Genome-Wide Analysis of the D-type Cyclin Gene Family Reveals Differential Expression Patterns and Stem Development in the Woody Plant Prunus mume

Cyclins, a prominent class of cell division regulators, play an extremely important role in plant growth and development. D-type cyclins (CYCDs) are the rate-limiting components of the G1 phase. In plants, studies of CYCDs are mainly concerned with herbaceous plants, yet little information is available about these genes in perennial woody plants, especially ornamental plants. Here, twelve Prunus mume CYCD (PmCYCDs) genes are identified and characterized. The PmCYCDs were named on the basis of orthologues in Arabidopsis thaliana and Oryza sativa. Gene structure and conserved domains of each subgroup CYCDs was similar to that of their orthologues in A. thaliana and O. sativa. However, PmCYCDs exhibited different tissue-specific expression patterns in root, stem, leaf, bud, and fruit organs. The results of qRT-PCR showed that all PmCYCDs, except PmCYCD5;2 and PmCYCD7;1, were primarily highly expressed in leaf buds, shoots, and stems. In addition, the transcript levels of PmCYCD genes were analyzed in roots under different treatments, including exogenous applications of NAA, 6-BA, GA3, ABA, and sucrose. Interestingly, although PmCYCDs were induced by sucrose, the extent of gene induction among PmCYCD subgroups varied. The induction of PmCYCD1;2 by hormones depended on the presence of sucrose. PmCYCD3;1 was stimulated by NAA, and induction was strengthened when sugar and hormones were applied together. Taken together, our study demonstrates that PmCYCDs are functional in plant stem development and provides a basis for selecting members of the cyclin gene family as candidate genes for ornamental plant breeding.