Petunia hybrida is a New Host of Pectobacterium brasiliense Associated with Soft Rot Disease in China.

Petunia hybrida is commonly cultivated for ornamental use in urban parks greening and street embellishment in China. In March 2022, 60% of P. hybrida plants cv. Wave Purple (n≈1800) from an ornamental plant nursery under natural conditions in Tianhe district (N 113°21'21", E 23°9'3.5"), Guangzhou, Guangdong Province, China, were affected with soft rot disease. The distribution of the disease was generally uniform. Infected plants initially exhibit small water-soaked lesions at the base of the stem, which then extended to the leaves. Eventually the diseased plant collapsed and died. Nine diseased plants were collected, and affected tissues cut into small fragments (5 × 5 mm), which were disinfested in 75% ethanol (30 s) and 2% sodium hypochlorite (60 s), followed by three rinses with sterile distilled water. The sterilized sections were macerated in 200 µl sterile water, and streaked on Luria-Bertani (LB) agar medium and incubated at 28°C for 48 h. Single colonies were restreaked three times to obtain purified isolation. Sixteen bacterial strains with similar morphology were isolated, and their colonies were yellowish white, round, and convex with smooth surfaces on LB agar plate. The representative strain BDQ1 was selected for further analyses and the 16S rDNA gene (GenBank Accession ON982467) were amplified using primer pair 27F/1492R, revealed above 99% sequence identity with some Pectobacterium brasiliense isolates (GenBank Accession Nos. CP046380(1421/1422), MN393966(1419/1422), and CP020350(1419/1422)) using BLASTn. A multilocus phylogenetic analysis by neighbor-joining method (1,000 bootstrap values) based on six housekeeping gene sequences of gyrA (GenBank Accession No. ON995454), icdA (ON995455), mdh (ON995456), mtlD (ON995457), proA (ON995458), and rpoS genes (ON995459) (Ma et al. 2007; Waleron et al., 2008). The results of phylogenetic analysis showed BDQ1 strain belong to the P. brasiliense clade. Pathogenicity tests were performed on ten healthy P. hybrida cv. Wave Purple plants by injecting 10 µl of bacterial suspensions of BDQ1 (108 CFU/ml) into the stems; another 10 healthy control plants were injected with 10 µl of sterile water. All plants were grown at 25-30°C and 60% humidity in natural light/dark cycle. After 3 d, all inoculated plants showed soft rot symptoms resembling to those observed in the nursery, while control plants remained healthy. Bacteria were successfully reisolated from the symptomatic tissues and identified to be P. brasiliense by PCR mentioned above. P. brasiliense is considered a very aggressive pathogen, which has been reported in Eurasia and Africa (Oulghazi et al. 2021). To our knowledge, this is the first report of P. brasiliense causing bacterial soft rot on P. hybrida in China. This pathogen may pose threat to P. hybrida production in area with warmand humid climate in China. The current study expands the known host range of P. brasiliense and helped raise attention on controlling pathogen spread.

Characterization and expression analysis of WRKY genes during leaf and corolla senescence of Petunia hybrida plants.

Several families of transcription factors (TFs) control the progression of senescence. Many key TFs belonging to the WRKY family have been described to play crucial roles in the regulation of leaf senescence, mainly in Arabidopsis thaliana. However, little is known about senescence-associated WRKY members in floricultural species. Delay of senescence in leaves and petals of Petunia hybrida, a worldwide ornamental crop are highly appreciated traits. In this work, starting from 28 differentially expressed WRKY genes of A. thaliana during the progression of leaf senescence, we identified the orthologous in P. hybrida and explored the expression profiles of 20 PhWRKY genes during the progression of natural (age-related) leaf and corolla senescence as well as in the corollas of flowers undergoing pollination-induced senescence. Simultaneous visualization showed consistent and similar expression profiles of PhWRKYs during natural leaf and corolla senescence, although weak expression changes were observed during pollination-induced senescence. Comparable expression trends between PhWRKYs and the corresponding genes of A. thaliana were observed during leaf senescence, although more divergence was found in petals of pollinated petunia flowers. Integration of expression data with phylogenetics, conserved motif and cis-regulatory element analyses were used to establish a list of candidates that could regulate more than one senescence process. Our results suggest that several members of the WRKY family of TFs are tightly linked to the regulation of senescence in P. hybrida. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01243-y.

Identification and expression analysis of NAC transcription factors potentially involved in leaf and petal senescence in Petunia hybrida.

Progression of leaf senescence depends on several families of transcription factors. In Arabidopsis, the NAC family plays crucial roles in the modulation of leaf senescence; however, the mechanisms involved in this NAC-mediated regulation have not been extensively explored in agronomic species. Petunia hybrida is an ornamental plant that is commonly found worldwide. Decreasing the rate of leaf and petal senescence in P. hybrida is essential for maintaining plant quality. In this study, we examined the NAC-mediated networks involved in regulating senescence in this species. From 41 NAC genes, the expression of which changed in Arabidopsis during leaf senescence, we identified 29 putative orthologs in P. hybrida. Analysis using quantitative real-time-PCR indicated that 24 genes in P. hybrida changed their transcript levels during natural leaf senescence. Leaf-expressed genes were subsequently assessed in petals undergoing natural and pollination-induced senescence. Expression data and phylogenetic analysis were used to generate a list of 10-15 candidate genes; 7 of these were considered key regulatory candidates in senescence because of their consistent upregulation in the three senescence processes examined. Altogether, we identified common and distinct patterns of gene expression at different stages of leaf and petal development and during progression of senescence. The results obtained in this study will contribute to the understanding of NAC-mediated regulatory networks in petunia.

Biochar versus hydrochar as growth media constituents for ornamental plant cultivation

Biochar and hydrochar have been proposed as novel materials for providing soilless growth media. However, much more knowledge is required before reliable advice can be given on the use of these materials for this purpose. Depending on the material and the technology applied (pyrolysis or hydrothermal carbonization), phytotoxicity and greenhouse gas emissions have been found for certain chars. In this study, our aim was to assess the feasibility of three chars as substrate constituents. We compared two biochars, one from forest waste and the other from olive mill waste, and a hydrochar from forest waste. We studied how chars affected substrate characteristics, plant performance, water economy and respiratory CO2 emission. Substrates containing biochar from forest waste showed the best characteristics, with good air/water relationships and adequate electrical conductivity. Those with biochar from olive mill waste were highly saline and, consequently, low quality. The substrates with hydrochar retained too much water and were poorly aerated, presenting high CO2 concentrations due to high respiratory activity. Plants performed well only when grown in substrates containing a maximum of 25 % biochar from forest waste or hydrochar. After analyzing the char characteristics, we concluded that biochar from forest waste could be safely used as a substrate constituent and is environmentally friendly when applied due to its low salinity and low CO2 emission. However, biochar from olive mill waste and hydrochar need to be improved before they can be used as substrate constituents.(AU)

Biochar versus hydrochar as growth media constituents for ornamental plant cultivation

ABSTRACT Biochar and hydrochar have been proposed as novel materials for providing soilless growth media. However, much more knowledge is required before reliable advice can be given on the use of these materials for this purpose. Depending on the material and the technology applied (pyrolysis or hydrothermal carbonization), phytotoxicity and greenhouse gas emissions have been found for certain chars. In this study, our aim was to assess the feasibility of three chars as substrate constituents. We compared two biochars, one from forest waste and the other from olive mill waste, and a hydrochar from forest waste. We studied how chars affected substrate characteristics, plant performance, water economy and respiratory CO2 emission. Substrates containing biochar from forest waste showed the best characteristics, with good air/water relationships and adequate electrical conductivity. Those with biochar from olive mill waste were highly saline and, consequently, low quality. The substrates with hydrochar retained too much water and were poorly aerated, presenting high CO2 concentrations due to high respiratory activity. Plants performed well only when grown in substrates containing a maximum of 25 % biochar from forest waste or hydrochar. After analyzing the char characteristics, we concluded that biochar from forest waste could be safely used as a substrate constituent and is environmentally friendly when applied due to its low salinity and low CO2 emission. However, biochar from olive mill waste and hydrochar need to be improved before they can be used as substrate constituents.

Biochar versus hydrochar as growth media constituents for ornamental plant cultivation

ABSTRACT Biochar and hydrochar have been proposed as novel materials for providing soilless growth media. However, much more knowledge is required before reliable advice can be given on the use of these materials for this purpose. Depending on the material and the technology applied (pyrolysis or hydrothermal carbonization), phytotoxicity and greenhouse gas emissions have been found for certain chars. In this study, our aim was to assess the feasibility of three chars as substrate constituents. We compared two biochars, one from forest waste and the other from olive mill waste, and a hydrochar from forest waste. We studied how chars affected substrate characteristics, plant performance, water economy and respiratory CO2 emission. Substrates containing biochar from forest waste showed the best characteristics, with good air/water relationships and adequate electrical conductivity. Those with biochar from olive mill waste were highly saline and, consequently, low quality. The substrates with hydrochar retained too much water and were poorly aerated, presenting high CO2 concentrations due to high respiratory activity. Plants performed well only when grown in substrates containing a maximum of 25 % biochar from forest waste or hydrochar. After analyzing the char characteristics, we concluded that biochar from forest waste could be safely used as a substrate constituent and is environmentally friendly when applied due to its low salinity and low CO2 emission. However, biochar from olive mill waste and hydrochar need to be improved before they can be used as substrate constituents.

Molecular insights into the purple-flowered ancestor of garden petunias.

PREMISE OF THE STUDY: The garden petunia is derived from Petunia axillaris (white flowered) and a purple-flowered species in the P. integrifolia group; it is still unclear which purple-flowered species was used in the initial crosses. This widely cultivated hybrid is an ideal model for different areas of scientific inquiry. METHODS: We analyzed three taxa of the P. integrifolia group considered to be the most probable parental candidates, along with a random sample of garden petunia representatives that were selected for their genetic variability. We used cpDNA trnH-psbA and trnS-trnG haplotypes and seven nuclear microsatellites in a population approach to investigate the genetic variability. This is the first time information from plastid DNA sequences and nuclear microsatellites has been combined to infer evolutionary relationships in these taxa. KEY RESULTS: Our results suggest that P. interior is the purple ancestor of garden petunias, and we postulate that the initial crosses must have been between both parents as a mother plant. CONCLUSIONS: Our work will contribute to the clarification of the evolutionary relationships among the Petunia ×hybrida and P. integrifolia taxa group and could be useful in breeding programs to transfer desired traits from wild to cultivated species.

Transgenic plants of Petunia hybrida harboring the CYP2E1 gene efficiently remove benzene and toluene pollutants and improve resistance to formaldehyde.

The CYP2E1 protein belongs to the P450 enzymes family and plays an important role in the metabolism of small molecular and organic pollutants. In this study we generated CYP2E1 transgenic plants of Petunia using Agrobacterium rhizogenes K599. PCR analysis confirmed that the regenerated plants contained the CYP2E1 transgene and the rolB gene of the Ri plasmid. Southern blotting revealed the presence of multiple copies of CYP2E1 in the genome of transgenic plants. Fluorescent quantitative PCR revealed exogenous CYP2E1 gene expression in CYP2E1 transgenic plants at various levels, whereas no like expression was detected in either GUS transgenic plants or wild-types. The absorption of benzene and toluene by transgenic plants was analyzed through quantitative gas chromatography. Transgenic plants with high CYP2E1 expression showed a significant increase in absorption capacity of environmental benzene and toluene, compared to control GUS transgenic and wild type plants. Furthermore, these plants also presented obvious improved resistance to formaldehyde. This study, besides being the first to reveal that the CYP2E1 gene enhances plant resistance to formaldehyde, also furnishes a new method for reducing pollutants, such as benzene, toluene and formaldehyde, by using transgenic flowering horticultural plants.

Transgenic plants of Petunia hybrida harboring the CYP2E1 gene efficiently remove benzene and toluene pollutants and improve resistance to formaldehyde

The CYP2E1 protein belongs to the P450 enzymes family and plays an important role in the metabolism of small molecular and organic pollutants. In this study we generated CYP2E1 transgenic plants of Petunia using Agrobacterium rhizogenes K599. PCR analysis confirmed that the regenerated plants contained the CYP2E1 transgene and the rolB gene of the Ri plasmid. Southern blotting revealed the presence of multiple copies of CYP2E1 in the genome of transgenic plants. Fluorescent quantitative PCR revealed exogenous CYP2E1 gene expression in CYP2E1 transgenic plants at various levels, whereas no like expression was detected in either GUS transgenic plants or wild-types. The absorption of benzene and toluene by transgenic plants was analyzed through quantitative gas chromatography. Transgenic plants with high CYP2E1 expression showed a significant increase in absorption capacity of environmental benzene and toluene, compared to control GUS transgenic and wild type plants. Furthermore, these plants also presented obvious improved resistance to formaldehyde. This study, besides being the first to reveal that the CYP2E1 gene enhances plant resistance to formaldehyde, also furnishes a new method for reducing pollutants, such as benzene, toluene and formaldehyde, by using transgenic flowering horticultural plants.