Structure of Petunia hybrida Defensin 1, a Novel Plant Defensin with Five Disulfide Bonds

The structure of a novel plant defensin isolated from the flowers of Petunia hybrida has been determined by H-1 NMR spectroscopy. P. hybrida defensin 1 (PhD1) is a basic, cysteine-rich, antifungal protein of 47 residues and is the first example of a new subclass of plant defensins with five disulfide bonds whose structure has been determined. PhD1 has the fold of the cysteine-stabilized alphabeta motif, consisting of an alpha-helix and a triple-stranded antiparallel beta-sheet, except that it contains a fifth disulfide bond from the first loop to the alpha-helix. The additional disulfide bond is accommodated in PhD1 without any alteration of its tertiary structure with respect to other plant defensins. Comparison of its structure with those of classic, four-disulfide defensins has allowed us to identify a previously unrecognized hydrogen bond network that is integral to structure stabilization in the family.

Methyl jasmonate vapour treatment suppresses specking caused by Botrytis cinerea on cut Freesia hybrida L. flowers

Treatment of cut freesia var. Cote d'Azur flowers with methyl jasmonate (MeJA, 0.1 mu l MeJA l(-1)) vapour suppressed petal specking caused by Botrytis cinerea infection. MeJA efficacy was concentration and incubation temperature dependent. Disease severity, lesion numbers and lesion diameters decreased with increasing MeJA concentration from 0.025 to 0.1 mu l MeJA l(-1). However, there were no significant (P > 0.05) differences among MeJA concentrations examined. MeJA was more effective in reducing B. cinerea flower specking at 20 degrees C than at 12 degrees C. MeJA treatment was ineffective at 5 degrees C. At 20 degrees C, MeJA treatment at 0.1 mu l MeJA l(-1) reduced disease severity, lesion numbers and lesion diameters by 58, 50 and 48%, respectively, as compared to untreated controls. In a repeat experiment, disease severity, lesion numbers and lesion diameters on MeJA vapour treated flowers after 12 h of incubation were reduced by 68, 56 and 50%, respectively. MeJA did not exert direct antifungal activity in-vitro, suggesting that treatment in-vivo reduced B. cinerea-induced flower specking by induction of host defence responses. MeJA at 0.1 mu l MeJA l(-1) significantly (P < 0.05) increased vase life of cut freesia flowers and delayed senescence judged by lower wilt scores and higher fresh weights as compared to untreated controls. (c) 2005 Elsevier B.V. All rights reserved.

Postharvest infection of Freesia hybrida flowers by Botrytis cinerea

'Specking' on harvested freesia (Freesia hybrida) flowers is a problem worldwide. The disease is caused by the fungal pathogen Botrytis cinerea. This disease symptom detracts from appearance and reduces marketability of the flowers. Unlike other important cut flower crops (e.g. gerbera), the mode of infection and epidemiology of postharvest freesia flower specking caused by B. cinerea has not been reported. Epidemiological studies were carried out under simulated conditions typical of those occurring during postharvest handling of freesia flowers. Infection of freesia flowers by B. cinerea occurred when a conidium germinated, formed a germ tube(s) and penetrated epidermal cells. Fungal hyphae then colonised adjacent cells, resulting in visible lesions. Different host reactions were observed on freesia 'Cote d'Azur' petals at 20 degrees C compared to 5 degrees C. The infection process was relatively rapid at 20 degrees C, with visible lesions produced within 7 h of incubation. However, lesion expansion ceased after 24 h of incubation. Infection was slower at 5 degrees C, with visible lesions produced after 48 h of incubation. However, lesion development at 5 degrees C was continuous, with lesions expanding over 4 days. Light microscopy observations revealed increased host defence reactions during infection. These reactions involved production of phenolic compounds, probably lignin and/or callose, around infection sites. Such substances may play a role in restricting petal colonisation and lesion expansion. Disease severity and lesion numbers on freesia flowers incubated at 12 degrees C were higher, but not significantly higher (P > 0.05), than on those incubated at 20 degrees C. Disease severity and progression were differentially mediated by temperature and relative humidity (R. H.). Infection of freesia flowers was severe at 100% R. H. for all three incubation temperatures of 5, 12 and 20 degrees C. In contrast, no lesions were produced at 80 to 90% R. H. at either 5 or 20 degrees C.

A survey of possible associations between preharvest environment conditions and postharvest rejections of cut freesia flowers

Botrytis cinerea is the major pathogen infecting cut freesia flowers. Flecking symptoms on petals caused by this fungus result in postharvest rejections and substantial economic loss to both growers and sellers. In a limited survey for industry, numbers of freesia stems sent from a specialist grower in The Netherlands and rejected at a cut flower wholesaler in the United Kingdom were documented. Relationships between preharvest environment conditions in Holland that may predispose flowers to infection and postharvest freesia rejection levels in the United Kingdom due to B. cinerea flecking symptom expression are reported. Freesia rejections peaked during spring and, to a lesser degree, autumn periods. However, no clear correlations between preharvest growing environment conditions (e.g. 3-day means for temperature preceding harvest) and postharvest rejection frequency (%) could be discerned. Thus, sporadic freesia rejections in the United Kingdom were probably attributable either to other unresolved variables during the pre- (e.g. infection pressure) and/or postharvest (e.g. condensation events) phases or to interactions among predisposing variables.

Effects of vase solution pH and abscisic acid on the longevity of cut 'Baccara' roses

Abscisic acid (ABA) supplied in the vase solution can induce stomatal closure in the leaves of cut flowers, including roses (Rosa hybrida L.). This effect may be beneficial in reducing water deficit stress. Extracellular pH can affect active ABA concentrations in the apoplast of guard cells, with sap alkalisation enhancing the physiological activity of ABA. Accordingly, it was hypothesized that vase solution pH may affect ABA-mediated stomatal closure of cut roses. Two experiments were conducted to study the interaction of vase solution pH and ABA. In the first, cut 'Baccara' roses were held in vase solutions with +/- 10(-5) M ABA at pH 6, pH 7 and pH 8. In the second experiment, roses were held with +/- 10(-5) M ABA at pH 6 and pH 8 in the presence and absence of 1 mg l(-1) AgNO3 as a bactericide. Supply of ABA increased vase life and reduced vase solution usage of flowers held in low pH 6 solutions, indicating induction of stomatal closure. Conversely, ABA supplied at pH 8 was associated with reduced vase life. This negative result was associated with enhanced development of vase solution microbes at high pH, which overrode any potential pH-mediated ABA efficacy effects.

Development and senescence of Grevillea 'Sylvia' inflorescences, flowers and flower parts

To characterise the physiology of development and senescence for Grevillea 'Sylvia'. oral organs, respiration, ethylene production and ACC concentrations in harvested flowers and flower parts were measured. The respiration rate of harvested inflorescences decreased over time during senescence. In contrast, both ethylene production and ACC concentration increased. Individual flowers, either detached from cut inflorescences held in vases at 20degreesC or detached from in planta inflorescences at various stages of development, had similar patterns of change in ACC concentration and rates of respiration and ethylene production as whole inflorescences. The correlation between ACC concentration and ethylene production by individual flowers detached from cut inflorescences held in vases was poor (r(2)=0.03). The isolated complete gynoecium (inclusive of the pedicel) produced increasing amounts of ethylene during development. Further sub-division of flower parts and measurement of their ethylene production at various stages of development revealed that the distal part of the gynoecium (inclusive of the stigma) had the highest rate of ethylene production. In turn, anthers had higher rates of ethylene production and also higher ACC concentrations than the proximal part of the gynoecium (inclusive of the ovary). Rates of ethylene production and ACC concentrations for tepal abscission zone tissue and adjacent central tepal zone tissue were similar. ACC concentration in pollen was similar to that in senescing perianth tissue. Overall, respiration, ethylene and ACC content measurements suggest that senescence of G. 'Sylvia' is non-climacteric in character. Nonetheless, the phytohormone ethylene is produced and evidently mediates normal flower development and non-climacteric senescence processes.

Effect of seasonal variation and storage temperature on leaf chlorophyll fluorescence and vase life of cut roses

Low temperature injury (LTI) of roses (Rosa hybrida L.) is difficult to assess by visual observation. Relative chlorophyll fluorescence (CF; F-v/F-m) is a non-invasive technique that provides an index of stress effects on photosystem 11 (PS 11) activity. This instrumental technique allows determination of the photosynthetic efficiency of plant tissues containing chloroplasts, such as rose leaves. In the present study, pre- and Post-Storage measurements of F-v/F-m were carried out to assess LTI in 'First Red' and 'Akito' roses harvested year round. Relationships between the pre-harvest environment conditions of temperature, relative humidity and photon flux density (PFD), F-v/F-m, and, vase life duration after storage are reported. After harvest, roses were stored at 1, 5 and 10 degrees C for 10 days. Non-stored roses were the control treatment. F-v/F-m ratios were reduced following storage, suggesting LTI of roses. However, reductions in F-v/F-m were not closely correlated with reduced vase life duration and were seasonally dependent. Only during winter experiments was F-v/F-m of roses stored at 1 degrees C significantly (P <= 0.001) lower compared to F-v/F-m of non-stored control roses and roses stored at 5 and 10 degrees C. Thus, the fall of F-v/F-m was due to an interaction of growing season and storage at 1 degrees C. Vase lives of roses grown during winter were significantly (P <= 0.001) shorter compared to roses grown during summer. Length of vase life was intermediate for roses grown during autumn and spring. Because of the lack of correlation between F-v/F-m and post-storage vase life it is concluded that the CF parameter F-v/F-m is nota practical index for assessing LTI in cold-stored roses. Higher PFD and temperature in summer were positively and significantly correlated with maintenance of post-storage FvIF ratios and longer vase life. It is suggested that shorter vase lives and lower post-storage F-v/F-m values after storage at 1 degrees C are consequences of reduced photosynthesis and smaller carbohydrate pools in winter-harvested roses. (c) 2004 Elsevier B.V All rights reserved.

Acibenzolar-S-methyl and methyl jasmonate treatments of glasshouse-grown freesias suppress post-harvest petal specking caused by Botrytis cinerea

Compounds that activate host plant defence responses potentially offer socio-environmentally sound alternative methods for disease control. In a series of glasshouse trials over 2 years, pre-harvest sprays with acibenzolar-S-methyl (ASM) and methyl jasmonate (MeJA) were tested for suppression of post-harvest infection of cut Freesia hybrida L. flowers by Botrytis cinerea. For the ASM treatments, variability in reducing the incidence of B. cinerea disease was observed between years freesia varieties, incubation temperatures and ASM concentrations. In the first year, the greatest reductions in lesion numbers on ASM-treated var. 'Cote d'Azur' were recorded using 2.86 mM ASM. For three different post-harvest temperature regimes, the relative reductions in lesion numbers, compared to untreated controls, were 45% at 5 degrees C, 40% at 12 degrees C and 30% at 20 degrees C, respectively. In the second year, lesion numbers were most reduced using 1.43 mM ASM to treat freesia var. 'Dukaat' flowers. Here, the relative reductions were to 44% at 5 degrees C, 26% at 12 degrees C and 51% at 20 degrees C. MeJA treatments were, in general, more consistently effective than ASM treatments in reducing lesion numbers and lesion diameters on cut freesia flowers. MeJA-treated (0.2 mM) freesia flowers (var. 'Dukaat') incubated at 20 degrees C showed relative reductions of 62%, and 45% for lesion number and lesion diameter, respectively. The differing efficacy between ASM and MeJA treatments could be attributed to their differential abilities to induce the salicylic acid (SA)-mediated vs. the jasmonic acid (JA)-mediated host defence pathways, respectively.