Effect of meta-topolin on leaf senescence and rooting in Pelargonium × hortorum cuttings

Pelargonium (Pelargonium x hortorum) is grown as potted or bedding plants for their colourful, showy flowers and scented foliage. Absence of senescence symptoms in the leaves of Pelargonium cuttings, their capacity to initiate roots and continued growth of initiated roots is an important quality attribute. The effects of postharvest treatments with meta-topolin (mT) and thidiazuron (TDZ) to ‘Katinka’ Pelargonium cuttings were investigated. Leaves treated with mT or TDZ for 5 d had higher leaf chlorophyll contents than untreated controls. Exposing cuttings to mT had no effect on the rooting proportion (%) and average root diameter. Similarly, 0.05 mM mT had no effect on number of roots per cutting. However, mT slightly reduced root length, root surface area and total volume of the roots. TDZ severely inhibited adventitious root formation, thus it reduced all the root parameters investigated. In conclusion, mT is very active in retarding leaf senescence, and combined with the observed ease of rooting of cuttings after mT treatment, this treatment is a suitable alternative to TDZ in delaying the onset of leaf yellowing in ornamental crops.     

Ethylene biosynthesis in apricot: Identification of a ripening-related 1-aminocyclopropane-1-carboxylic acid synthase (ACS) gene

Apricots are climacteric fruits with a high susceptibility to flesh softening and loss of flavor during postharvest storage, and most of the ripening processes are regulated by ethylene, which also has an effect on its own biosynthesis. To understand this process in apricot, inhibition of ethylene biosynthesis and perception was performed for studying key genes involved in the ethylene biosynthetic pathway. Apricots, cv. “Patterson”, were harvested with yellow-green ground color and immediately treated with either the ethylene perception inhibitor 1-methyl cyclopropene (1-MCP) at 10 μL L⁻¹ or the ethylene biosynthesis inhibitor aminoethoxyvinylglycine (AVG) at 1 g L⁻¹. After treatment, quality and physiological attributes such as firmness, color, total soluble solids, acidity, fruit weight, ethylene production and respiration rates were evaluated every 2 d until they ripened at 20 °C. Gene expression analysis was performed by quantitative polymerase chain reaction (qPCR). Both ethylene inhibitors were effective in reducing ethylene production, respiration rate and fruit softening. Three 1-aminocyclopropane-1-carboxylic-acid synthase (ACS) genes were characterized, but only the expression of ACS2 was highly reduced by ethylene inhibition, suggesting a key role in ethylene synthesis at ripening. Contrarily, ACS1 and ACS3 showed a higher expression under ethylene inhibition suggesting that the corresponding genes are individually regulated in a specific mode as observed in other climacteric fruits. Finally, changes in 1-aminocyclopropane-1-carboxylic-acid oxidase genes did not show a consistent pattern of ethylene modulation.     

Molecular analysis of softening and ethylene synthesis and signaling pathways in a non-softening apple cultivar, ‘Honeycrisp’ and a rapidly softening cultivar, ‘McIntosh’

A feature of ‘Honeycrisp’ apples [Malus sylvestris (L.) Mill var. domestica (Borkh.) Mansf.] is that they maintain flesh firmness over extended storage. The objective of this study was to elucidate molecular mechanisms that are responsible for slow softening of ‘Honeycrisp’ as compared with a rapidly softening cultivar, ‘McIntosh’. Fruit from both cultivars were picked during the normal harvest period and stored at 20 °C for 10 d. Internal ethylene concentrations (IECs) in ‘Honeycrisp’ fruit were lower than in ‘McIntosh’, but at climacteric levels of ethylene ‘Honeycrisp’ fruit maintained their firmness over this period, while ‘McIntosh’ softened rapidly. Concentrations of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) were higher in ‘Honeycrisp’ than in ‘McIntosh’ apples. qRT-PCR analysis was carried out for genes involved in ethylene biosynthesis, perception and signaling [ACC synthase (MdACS); ACC oxidase (MdACO); ethylene receptors (MdETR and MdERS); constitutive triple response (MdCTR); ethylene response factor (MdERF)], as well as those involved in cell wall metabolism [polygalacturonase (MdPG); xyloglucan endotransglucosylase (MdXTH); expansin (MdEXP); β-galactosidase (Md β-GS); arabinofuranosidase (MdAFase); pectate lyase (MdPL)]. At comparable IECs, the expression of genes involved in ethylene synthesis, ethylene perception and signal transduction was generally much higher in ‘Honeycrisp’ than in ‘McIntosh’ fruit. However, the expression of MdAFase and MdEXP3 was generally lower in ‘Honeycrisp’ than in ‘McIntosh’, while that of MdPG and MdPL was extremely low in ‘Honeycrisp’. Expression of MdPG1 was very low, even though IECs were at climacteric levels. Absence of fruit softening in ‘Honeycrisp’ is probably associated with restricted cell wall enzyme activity. The lower maximum IECs found in ‘Honeycrisp’ compared with ‘McIntosh’ do not appear to be related to expression of genes involved in ethylene biosynthesis.     

Effect of 1-methylcyclopropene (1-MCP) on reducing postharvest decay in tomatoes (Solanum lycopersicum L.)

1-Methylcyclopropene (1-MCP as SmartFresh™ technology), an ethylene antagonist, was evaluated for affecting postharvest decay caused by Alternaria alternata, Botrytis cinerea, and Fusarium spp. on ‘Quality 23’ and ‘Seminis 35’ tomatoes at green or pink stages. Fruit with natural or artificial infection were subjected to 1-MCP at 0.0 μL L⁻¹, 0.6 μL L⁻¹ for 12 h, and 1.0 μL L⁻¹ for 6 h. After 31-42 d storage, disease incidence and severity of individual diseases in 1-MCP treated fruit was significantly reduced compared with that of the untreated controls, except in one inoculated test for ‘Quality 23’ where severity of Alternaria rot in 1.0 μL L⁻¹ treated fruit were significantly higher than that of the untreated control. Fruit treated with 1-MCP at 1.0 L⁻¹ for 6 h also had significantly higher incidence of Alternaria rot in the inoculated ‘Quality 23’ and in the non-inoculated ‘Seminis 35’ compared with the fruit treated with 1-MCP at 0.6 μL L⁻¹ for 12 h. The results of this study indicate that 1-MCP can reduce postharvest decay within a certain storage period.     

蛋白激发子基因pemG1转化三生烟中及其对TMV抗性的提高

通过根癌农杆菌(Agrobactrium tumefaciens)介导转化法,将含有稻瘟菌蛋白激发子基因pemG1的植物表达载体pCAMBIA2300-Ubi-pemG1-Ocs转化三生烟(Nicotiana tobacum cv. Samsun NN),获得了转基因植株。用PCR检测抗卡那霉素烟苗确认阳性转化株,用Southern、Northern和Western杂交进一步证实了pemG1基因的整合、转录和表达。对T2代转基因阳性株进行烟草花叶病毒(Tobacco Mosaic Virus)接种试验。接种5 d后发现,与非转基因对照相比,表达pemG1的烟草叶片枯斑数量减少,表明稻瘟菌蛋白激发子基因pemG1的表达提高了转基因烟草对TMV的抗性。     

水稻(Oryza sativa)新型液泡Na+/H+逆向转运蛋白基因的特征分析和表达

以拟南芥AtNHX1 cDNA 片段作为探针，筛查水稻盐胁迫植株叶片cDNA 文库，获得与AtNHX1同源的水稻新型液泡Na⁺/H⁺逆向转运蛋白基因（OsANT1）。序列分析表明，OsANT1 全长cDNA为2 178 bp，包括一个长度为1 608 bp的完整开放阅读框，编码535个氨基酸残基。在DNA水平上，OsANT1基因含有15个外显子和14个内含子，长度为4 835 bp。OsANT1含有12个跨膜域，系统进化树分析结果表明，与来自拟南芥、水稻、小麦、玉米、大麦、马蔺和芦苇等的Na⁺/H⁺逆向转运蛋白高度同源。盐胁迫条件下，OsANT1的表达具有盐分诱导特征，且随着胁迫的增大而增加。表明该基因可能在水稻抵御盐分胁迫的过程中具有一定作用。     

Evaluation of the role of the endo-β-(1,4)-glucanase gene FaEG3 in strawberry fruit softening

In strawberry, Fragaria × ananassa Duch., fruit, two different endo-β-1,4-glucanase genes, FaEG1 and FaEG3, also named Cel1 and Cel2, are expressed during the softening process that occurs during fruit ripening. It has also been suggested that FaEG3, which contains a putative cellulose-binding domain, could play a key role in fruit development, since previous attempts to down-regulate this gene through transgenesis have been unsuccessful. In this investigation, we obtained transgenic strawberry plants containing an antisense sequence of the FaEG3 gene under the control of the 35S promoter. Ripened fruit from transgenic lines (Acel lines) showed large variation in FaEG3 silencing, but fruit firmness was similar to control fruit in all the lines. Two Acel lines showing almost 95% reductions in FaEG3 mRNA levels were selected for further study. In these lines, FaEG3 down-regulation was high, from 78 to 95%, at all fruit developmental stages, whereas FaEG1 was only slightly suppressed. In spite of the high FaEG3 silencing achieved, EGase activity was not modified in ripe fruit. At the cell wall level, walls from transgenic ripe fruit contained a significantly higher amount of the 4 M KOH fraction, which is enriched in hemicellulosic polymers. The analysis of this fraction by size exclusion chromatography showed that transgenic cell walls contained a smaller amount of higher molecular mass polymers than controls. Altogether, these results indicate that FaEG3 does not play a key role either in fruit development or fruit softening. However, its silencing affects the amount and, in a minor way, the size of hemicellulosic polymers.