Mechanisms generating the numerous hues of chrysanthemum petals can be traced to in vitro γ-ray-induced mutagenesis.

In terms of global cut flower production, chrysanthemum (Dendranthemum grandiflorum kitam.) ranks among the top ten. When a plant species possesses colour characteristics associated with those of ancestral species, that colour is described as being white, yellow, or pink, with white coloration representing an absence of pigments, and yellow and pink coloration each represents the presence of carotenoids and anthocyanins, respectively. Pigments, or a combination of pigments, have been improved upon to provide an expansive rainbow of floral colours like orange, dark red, purplish-red, and red. Recent green-flowered varieties have developed a reputation for containing chlorophylls in their ray petals. Additionally, transgenic interventions have been used to breed violet/blue flowers. Flower colour is a highly regarded characteristic of any flower cultivar, particularly chrysanthemum. Breeders' ultimate goal is to create newer chrysanthemum cultivars with innovative characteristics, such as new flower colours, using a time and input-efficient approach. Investigating the molecular mechanisms that regulate flower pigmentation may yield critical insights into the rational manipulation of floral colour. To generate a diverse array of flower colour mutants in chrysanthemum cv. “Candid” through mutagenesis, in vitro grown micro shoots were exposed to 10, 20, 30, and 40 Gy gamma irradiation at 100 Gy per minute and were evaluated for different parameters. The rhizogenesis parameters declined with the increase in irradiation dose from 0 Gy to 40 Gy, while as, 10 Gy dose proved to record minimum decline in contrast to the control. Survival, leaf size, and the number of leaves plant-1 after the 8th-week interval exhibited a downward trend with the increasing trend of gamma irradiation dose but recorded a least drop in plants raised from shoots irradiated with 10 Gy gamma irradiation dose with respect to the control. Apparently, the minimum delay in the number of days to floral bud appearance took under 10 Gy compared to control. The highest number of flower colour mutants was recorded under 10 Gy (light pink, orange-pink, white and yellow). Demountable mutation frequency based on flower colour was desirable in plants irradiated with the slightest dose of 10 Gy.

Combination Breeding and Marker-Assisted Selection to Develop Late Blight Resistant Potato Cultivars

(1) Background: Although resistance to pathogens and pests has been researched in many potato cultivars and breeding lines with DNA markers, there is scarce evidence as to the efficiency of the marker-assisted selection (MAS) for these traits when applied at the early stages of breeding. A goal of this study was to estimate the potential of affordable DNA markers to track resistance genes that are effective against the pathogen Phytophthora infestans (Rpi genes), as a practical breeding tool on a progeny of 68 clones derived from a cross between the cultivar Sudarynya and the hybrid 13/11-09. (2) Methods: this population was studied for four years to elucidate the distribution of late blight (LB) resistance and other agronomical desirable or simple to phenotype traits such as tuber and flower pigmentation, yield capacity and structure. LB resistance was phenotypically evaluated following natural and artificial infection and the presence/absence of nine Rpi genes was assessed with 11 sequence-characterized amplified region (SCAR) markers. To validate this analysis, the profile of Rpi genes in the 13/11-09 parent was established using diagnostic resistance gene enrichment sequencing (dRenSeq) as a gold standard. (3) Results: at the early stages of a breeding program, when screening the segregation of F1 offspring, MAS can halve the workload and selected SCAR markers for Rpi-genes provide useful tools.

Combination Breeding and Marker-Assisted Selection to Develop Late Blight Resistant Potato Cultivars

(1) Background: Although resistance to pathogens and pests has been researched in many potato cultivars and breeding lines with DNA markers, there is scarce evidence as to the efficiency of the marker-assisted selection (MAS) for these traits when applied at the early stages of breeding. A goal of this study was to estimate the potential of affordable DNA markers to track Rpi disease resistance genes, that are effective against the pathogen Phytophthora infestans, as a practical breeding tool on a progeny of 68 clones derived from a cross between the cultivar Sudarynya and 13/11-09. (2) Methods: this population was studied for four years to elucidate the distribution of LB resistance and other agronomical desirable or simple to phenotype traits such as tuber and flower pigmentation, capacity and structure of yield. LB resistance was phenotypically determined through natural and artificial infection and the presence/absence of nine Rpi genes was assessed via 11 sequence-characterized amplified region (SCAR) markers. To aid this analysis, the profile of Rpi genes in the 13/11-09 parent was established using diagnostic resistance gene enrichment sequencing (dRenSeq) as a gold standard. (3) Results: at the early stages of a breeding program, MAS can halve the workload when screening the segregation of F1 offspring and selected SCAR markers for Rpi-genes provide useful tools.

Congenial in Vitro γ-Ray Induced Mutagenesis Underlying the Varied Array of Petal Colours in Chrysanthemum (Dendranthemum Grandiflorum Kitam). ‘Candid’

Chrysanthemum (Dendranthemum grandiflorum kitam.) is amongst the top ten cut flowers globally. The flower colour of ancestral species is restricted to white, yellow, and pink and is acquired from anthocyanins, carotenoids, and the dearth of both pigments, respectively. An extensive array of flower colours, like orange, dark red, purplish-red, and red, has been bred by enhancing the variety of pigments or the amalgamation of both pigments. In recent times, green-flowered cultivars having chlorophylls in their ray petals have been produced and have grown a reputation. Furthermore, violet /blue flowers have been bred via transgenic interventions. Flower colour is considered as critically acclaimed feature of any flower cultivar especially chrysanthemum. Creating newer chrysanthemum cultivars with novel features, for instance, new flower colours in a time and input optimised approach, is the eventual ambition for breeders. Exploring the molecular mechanisms that control flower pigmentation may present imperative suggestions for the rational manoeuvring of flower colour. To generate a diverse array of flower colour mutants in chrysanthemum cv. “Candid” through mutagenesis, in vitro grown micro shoots were exposed to 10, 20, 30, and 40 Gy gamma irradiation at 100 Gy per minute and were evaluated for different parameters. The rhizogenesis parameters declined with the increase in irradiation dose from 0 Gy to 40 Gy, while as, 10 Gy dose proved to record minimum decline in contrast to the control. Survival, leaf size, and the number of leaves plant− 1 after the 8th -week interval also decreased with the increasing trend of gamma irradiation dose but recorded a minimum decline in plants raised from shoots irradiated with 10 Gy gamma irradiation dose with respect to the control. Apparently, the minimum delay in the number of days to floral bud appearance took under 10 Gy compared to control. The highest number of flower colour mutants were recorded under 10 Gy (light pink, orange-pink, white and yellow). Demountable mutation frequency based on flower colour was desirable in plants irradiated with the slightest dose of 10 Gy

The Conserved and Particular Roles of the R2R3-MYB Regulator FhPAP1 from Freesia hybrida in Flower Anthocyanin Biosynthesis

Anthocyanin biosynthesis is mainly controlled by MYB–bHLH–WD40 (MBW) complexes that modulate the expression of anthocyanin biosynthetic genes (ABGs). The MYB regulators involved in anthocyanin biosynthesis arose early during plant evolution and thus might function divergently in different evolutionary lineages. Although the anthocyanin-promoting R2R3-MYB regulators in eudicots have been comprehensively explored, little consensus has been reached about functional discrepancies versus conservation among MYB regulators from different plant lineages. Here, we integrated transcriptome analysis, gene expression profiles, gain-of-function experiments and transient protoplast transfection assays to functionally characterize the monocot Freesia hybrida anthocyanin MYB regulator gene FhPAP1, which showed correlations with late ABGs. FhPAP1 could activate ABGs as well as TT8-clade genes FhTT8L, AtTT8 and NtAN1 when overexpressed in Freesia, Arabidopsis and tobacco, respectively. Consistently, FhPAP1 could interact with FhTT8L and FhTTG1 to form the conserved MBW complex and shared similar target genes with its orthologs from Arabidopsis. Most prominently, FhPAP1 displayed higher transactivation capacity than its homologs in Arabidopsis and tobacco, which was instantiated in its powerful regulation on ABGs. Moreover, we found that FhPAP1 might be the selected gene during the domestication and rapid evolution of the wild Freesia species to generate intensive flower pigmentation. These results showed that while the MBW complex was highly evolutionarily conserved between tested monocot and core eudicot plants, participating MYB regulators showed functional differences in transactivation capacity according to their activation domain and played important roles in the flower coloration domestication and evolution of angiosperms.

Identification of MicroRNAs and Their Targets Involved in Paeonia rockii Petal Variegation Using High-throughput Sequencing

Tree peony (Paeonia sp.) is a popular traditional ornamental plant in China. Among the nine wild species, Paeonia rockii displays wide-ranging, deep purple variegation at the base of the petals, whereas Paeonia ostii exhibits purely white petals. Overall, the posttranscriptional regulation involved in tree peony flower opening and pigmentation remains unclear. To identify potential microRNAs (miRNAs) involved in flower variegation, six small RNA libraries of P. ostii and P. rockii petals at three different opening stages were constructed and sequenced. Using Illumina-based sequencing, 22 conserved miRNAs and 27 novel miRNAs were identified in P. rockii and P. ostii petals. Seventeen miRNAs were differentially expressed during flower development, and several putative target genes of these miRNAs belonged to transcription factor families, such as Myb domain (MYB), and basic helix-loop-helix (bHLH) transcription factors. Furthermore, an integrative analysis of the expression profiles of miRNAs and their corresponding target genes revealed that variegation formation might be regulated by miR159c, miR168, miR396a, and novel_miR_05, which target the MYB transcription factors, chalcone synthase (CHS), and ABC transporter. Our preliminary study is the first report of miRNAs involved in Paeonia flower pigmentation. It provides insight regarding the molecular mechanisms underlying the regulation of flower pigmentation in tree peony.