Regulation of stony hard peach softening with ACC treatment

Controlling the rate of fruit softening in melting-flesh peaches is a primary goal of the fruit industry. Stony hard (SH) peach varieties lack the ability to synthesize 1-aminocyclopropane-1-carboxylic acid (ACC), the precursor of ethylene, which is required for fruit maturation. SH peaches thus have crisp flesh that remains firm during ripening. In this study, we developed a simple technique to stimulate fruit softening by a single spray application of ACC at a concentration of 10–20 mM, which was sufficient to allow ethylene synthesis and fruit softening. Higher concentrations of ACC increased ethylene production, and made the fruit softer. Ethylene synthesis was limited to the first 2–3 d after ACC treatment, after which fruit ceased softening and retained its remaining firmness. These results indicate that a single application of ACC solution can be used to regulate the process of fruit softening in SH peaches.     

Control of ripening in transgenic tomatoes

Major progress has been made over the last few years in the identification and regulation of tomato ripening genes. At least 25 genes showing elevated expression during ripening have been cloned and several, including polygalacturonase, which modifies fruit textures, have been shown to be ripening-specific. In addition, genes have been cloned for ACC synthase and ACC oxidase, which control the synthesis of ethylene, which plays a critical role in ripening. Inhibition of expression of polygalacturonase, pectinesterase, ACC synthase, ACC oxidase and phytoene synthase has been achieved in transgenic plants, using antisense technology. The expression of several genes has also been inhibited by sense gene suppression. New traits caused by these transgenes are stably inherited. Antisense tomatoes with reduced polygalacturonase have improved textural qualities which are being exploited commercially for the fresh and processed markets. Overexpression of phytoene synthase has been shown to restore carotenoid production in the yellow flesh mutant and can be used to enhance colour in other cultivars. Antisense fruit in which ACC synthase or ACC oxidase are inhibited show slower ripening and reduced over-ripening. ACC oxidase antisense genes have also been shown to delay leaf senescence. It is to be expected that further genes determining other quality traits will be identified and manipulated soon.     

1-MCP extends the storage and shelf life of mangosteen (Garcinia mangostana L.) fruit

Mangosteen (Garcinia mangostana L.) fruit were harvested when the peel (pericarp) was light greenish yellow with scattered pinkish spots. Fruit were exposed to 1 μL L⁻¹ 1-methylcyclopropene (1-MCP) for 6 h at 25 °C and were then stored at 25 °C (control) or 15 °C. The 1-MCP treatment only temporarily delayed softening of the fruit flesh, during storage. Storage life, defined as the time until the pericarp was dark purple, was much longer in fruit stored at 15 °C than in fruit stored at 25 °C. It was also longer in 1-MCP treated fruit (storage life at 15 °C: control 18 d, 1-MCP-treated fruit 27 d). The 1-MCP treatment also increased the length of shelf life, defined as the time until the pericarp turned blackish purple or showed calyx wilting, at 25 °C. 1-MCP treatment reduced ethylene production. It also reduced pericarp levels of 1-aminocyclopropane-1-carboxylic acid (ACC), and the pericarp activities of ACC synthase (ACS) and ACC oxidase (ACO). In the fruit flesh, in contrast, 1-MCP did not affect ACC levels and ACS activity, but the treatment reduced ACO activity. Taken together, both the storage life and the shelf life of the fruit were extended by the 1-MCP treatment. A decrease in ACO activity largely accounted for the effects of the 1-MCP on ethylene production in the pericarp.     

Morphological characterisation and agronomic evaluation of transgenic broccoli (Brassica oleracea L. var. italica) containing an antisense ACC oxidase gene

Morphological characterisation and agronomic evaluation was conducted on 12 transgenic broccolilines containing a tomato antisense1-aminocyclopropane-1-carboxylic acid (ACC) oxidase gene. Plants of three cultivars: Shogun (Sh), Green Beauty (Gy) and Dominator (D), were regenerated from hairy root cultures after co-cultivation with Agrobacterium rhizogenes strain A4T harbouring the binary vector pLN35. The T-DNA of pLN35 contains genes encoding a tomato antisense ACC oxidase gene (35S-ACC-5′7′) and a neomycin phosphotransferase II gene (NOS-NPTII-NOS) for kanamycin resistance. The transgenic plants were transferred to a greenhouse and fertile plants obtained. Integration of the foreign DNA into the broccoli genome was confirmed by the polymerase chain reaction and Southern analyses. Transgenic plants showed evidence of hairy root (HR)-induced morphological changes to varying degrees. Of the 12 characterised transgenic lines, three lines(Gy/7, D/1 and D/2) performed within the limits of acceptability for all head quality parameters analysed (size, density, colour, shape and leafiness). The ethylene production from stalks of four field-grown transgenic lines of Green Beauty broccoli showed significant reductions in activity relative to the control 98 h after harvest. The Dominator transgenic lines D/1 and D/2 showed significant improvements in head colour relative to the control from 48 h after harvest. These results are consistent with the ethylene production patterns determined previously for these lines. The head colour results are consistent with previous results suggesting that two enzyme systems may be involved in broccoli senescence, giving two bursts of ethylene production, with only the second burst inhibited by the antisense ACC oxidase gene used. This revised version was published online in August 2006 with corrections to the Cover Date.     

Regulation of propylene-induced ripening and ethylene biosynthesis by oxygen in ‘Hayward’ kiwifruit

Kiwifruit, cv Hayward, were treated with 130 μl/l propylene under 1, 5, 10, 13, 16, and 21% O₂ concentrations in a continuous flow-through system at 20 °C. Control kiwifruit were kept in air (21%O₂). Flesh firmness, soluble solids content, internal ethylene concentration, l-aminocyclopropane-l-carboxylic acid concentration (ACC), malonyl-l-aminocyclopropane-l-carboxylic acid concentration (MACC), and EFE capacity were measured during the experiment. Ethylene biosynthesis and fruit ripening were stimulated when kiwifruit were exposed to a mixture of propylene in 21% O₂. Reduction of oxygen concentration (< 10%) inhibited the effectiveness of propylene on autocatalytic ethylene production possibly through a reduction in ACC concentration in the tissue. Exposure of kiwifruit to low oxygen concentrations (< 5%) delayed propylene-induced fruit ripening.     

The manipulation and modification of tomato fruit ripening by expression of antisense RNA in transgenic plants

Summary The common cultivated tomato (Lycopersicon esculentum Mill.) provides a major focus for improvement of crop quality through genetic engineering. Identification of ripening-related cDNAs has enabled the modification of specific aspects of ripening by manipulating gene expression in transgenic plants. By utilizing antisense RNA to modify expression of ripening genes, we have inhibited the production of the cell wall-metabolising enzymes polygalacturonase and pectinesterase and created transgenic plants that contain, effectively, single, targeted mutations affecting these genes. Furthermore, this approach has been used with previously unidentified cDNA clones to enable both functional identification and manipulation of genes involved in ethylene production (ACC oxidase) and carotenoid biosynthesis (phytoene synthase). The use of antisense RNA targeted to specific genes to alter ripening phenotypes and improve commercial utility of fruit by affecting shelf-life, processing characteristics and nutritional content is discussed.We have used the extreme ripening-impaired mutant, ripening inhibitor (rin) to identify additional genes implicated in the ripening process. This approach has resulted in the cloning of several novel ripening-related mRNAs which are now being studied by antisense experiments. This may enable identification and manipulation of additional genes involved in processes such as softening, flavour and aroma generation and susceptibility to pathogens.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - PE pectinesterase - PG polygalacturonase - SAM S-adenosyl methionine - SARs scaffold attachment regions     

Mode of action of nitric oxide in inhibiting ethylene biosynthesis and fruit softening during ripening and cool storage of ‘Kensington Pride’ mango

The mode of action of nitric oxide (NO) in inhibiting ethylene biosynthesis and fruit softening during ripening and cool storage of mango fruit was investigated. Hard mature green mango (Mangifera indica L. cv. ‘Kensington Pride’) fruit were fumigated with 20 μL L⁻¹ NO for 2 h at 21 °C and allowed to ripen at 21 ± 1 °C for 10 d, or stored at 13 ± 1 °C for 21 d. During ripening and cool storage, ethylene production and respiration rate from whole fruit were determined daily. The 1-aminocyclopropane-1-carboxylic acid (ACC) content, activities of ACC synthase (ACS), ACC oxidase (ACO), and fruit softening enzymes such as pectin esterase (PE), endo-1,4-β-d-glucanase (EGase), exo- and endo-polygalacturonase (exo-PG, endo-PG) as well as firmness and rheological properties of pulp were determined at two- and seven-day intervals during ripening and cool storage, respectively. NO fumigation inhibited ethylene biosynthesis and respiration rate, and maintained higher pulp firmness, springiness, cohesiveness, chewiness, adhesiveness, and stiffness. NO-fumigated fruit during cool storage and ripening had lower ACC contents through inhibiting the activities of both ACS and ACO in the fruit pulp. NO-fumigated fruit showed decreased activities of exo-PG, endo-PG, EGase, but maintained higher PE activity in pulp tissues during ripening and cool storage. In conclusion, NO fumigation inhibited ethylene biosynthesis through inhibition of ACS and ACO activities leading to reduced ACC content in the fruit pulp which consequently, reduced the activities of fruit softening enzymes during ripening and cool storage.     

Effect of high temperature stress on ethylene biosynthesis, respiration and ripening of ‘Hayward’ kiwifruit

Temperatures up to 35°C have been shown to increase ethylene production and ripening of propylene-treated kiwifruit (Stavroulakis, G., Sfakiotakis, E.M., 1993. We attempted to study the regulation by high stress temperature of the propylene induced ethylene biosynthesis and ripening in ‘Hayward’ kiwifruit. ‘Hayward’ kiwifruit were treated with 130 μl/l propylene at temperatures from 30 to 45°C up to 120 h. Ethylene biosynthesis pathway and fruit ripening were investigated. Propylene induced normal ripening of kiwifruit at 30–34°C. Fruit failed to ripe normally at 38°C and above 40°C ripening was inhibited. Propylene induced autocatalytic ethylene production after a lag period of 24 h at 30–34°C. Ethylene production was drastically reduced at 38°C and almost nil at 40°C. The 1-aminocyclopropane-1-carboxylic acid (ACC) content was similar at 30–38°C and was very low at 40°C. The 1-aminocyclopropane-1-carboxylate synthase (ACC synthase) and 1-aminocyclopropane-1-carboxylate oxidase (ACC oxidase) activities decreased with a temperature increase above 30°C, but ACC oxidase decreased at a faster rate than ACC synthase. Fruit not treated with propylene showed no ripening response or ethylene production. However, kiwifruit respiration rate increased with temperature up to 45°C, reaching the respiration peak in 10 h. At temperatures up to 38°C, propylene treatment enhanced the respiration rate. After 48 h at 45°C, fruit showed injury symptoms and a larger decrease in CO₂. The results suggest that high temperature stress inhibits ripening by inhibiting ethylene production and sensitivity while respiration proceeds until the breakdown of tissues.     

Apparent synergism between the positive effects of 1-MCP and modified atmosphere on storage life of banana fruit

Fruit of cv. Gros Michel banana were treated with 1-MCP (1000 nL L⁻¹ for 4 h at 25 °C) and then packed in non-perforated polyethylene (PE) bags for modified atmosphere storage (MAP). The bags were placed in corrugated cardboard boxes and stored at 14 °C. Fruit were removed from cool storage and ripened at room temperature using ethephon. The length of storage life was determined by the change in peel color to yellow, after this ethephon treatment. Fruit treated with 1-MCP + MAP had a storage life of 100 days. The storage life of control fruit (no 1-MCP and no MAP) was 20 days. Fruit held in PE bags without 1-MCP treatment had a 40 day storage life, and the same was found in fruit treated with 1-MCP but without PE bags. 1-MCP is an inhibitor of ethylene action, but also inhibited ethylene production, mainly through inhibition of ACC oxidase activity in the peel. MAP inhibited ethylene production mainly through inhibition of ACC oxidase, both in the peel and pulp. The combination of 1-MCP treatment and MAP storage resulted in much lower ethylene production due to inhibition of both ACC synthase and ACC oxidase activity.     

桃果实中ACC合酶基因克隆及基因沉默载体构建

摘 要：植物中乙烯是一种具有促进果实成熟和衰老的内源激素。ACC合酶是植物乙烯生物合成途径中一个重要的限速酶,沉默ACC合酶基因的表达能减少植物性内源性乙烯的产生。本研究以中华寿桃为研究材料,采用RT-PCR 技术,克隆获得ACC合酶基因。将该基因酶切回收后连接到pTRV-RNA2载体上,转化DH5α,筛选阳性克隆,进行酶切鉴定。测序后与已知序列进行同源性比较,其同源性达到99.6%,表明将ACC合酶基因成功连接到pTRV-RNA2基因沉默载体上。     

Regulation of stony hard peach softening with ACC treatment

Controlling the rate of fruit softening in melting-flesh peaches is a primary goal of the fruit industry. Stony hard (SH) peach varieties lack the ability to synthesize 1-aminocyclopropane-1-carboxylic acid (ACC), the precursor of ethylene, which is required for fruit maturation. SH peaches thus have crisp flesh that remains firm during ripening. In this study, we developed a simple technique to stimulate fruit softening by a single spray application of ACC at a concentration of 10–20 mM, which was sufficient to allow ethylene synthesis and fruit softening. Higher concentrations of ACC increased ethylene production, and made the fruit softer. Ethylene synthesis was limited to the first 2–3 d after ACC treatment, after which fruit ceased softening and retained its remaining firmness. These results indicate that a single application of ACC solution can be used to regulate the process of fruit softening in SH peaches.     

高等植物ACC合成酶基因的克隆及其表达调控的研究进展

主要对高等植物ACC合成酶基因的克隆、结构及其表达调控等方面研究进展进行综述,旨在揭示ACC合成酶基因的分子特征,为运用基因工程技术探索控制果实成熟、软化和衰老的关键ACC合成酶基因,更进一步地通过转基因技术调控果实内乙烯的含量从而延长水果货架期提供思路。     

甜瓜ACS基因家族成员的鉴定及其表达特性分析

乙烯在高等植物生长发育等多个方面都具有重要的调控作用,而ACS(1-aminocyclopropane-1-carboxylic acid(ACC)synthase,1-氨基环丙烷-1羧酸合成酶)是乙烯生物合成途径的限速酶,由多基因家族编码,不同的发育、环境、激素刺激等信号诱导不同的ACS基因表达.本研究以甜瓜品种'河...     

西瓜数量性状遗传距离及杂种优势分析

为研究西瓜数量性状的遗传距离和杂种优势，对8份西瓜亲本采用NC-Ⅱ设计配制16个杂交组合，对杂交组合的7个数量性状进行相关性分析、主成分分析和杂种优势研究。结果表明，坐果节位和单果重显著正相关，中心可溶固形物含量和边部可溶固形物含量极显著正相关。主蔓长度、坐果节位、单果重、果皮厚度、中心可溶固形物含量、边部可溶固形物含量和果形指数7个数量性状可以综合成为3个主成分，且3个主成分构成的信息量可以解释总信息量的87.3132%。8个亲本间遗传距离介于0.3315~1.5121之间，A02×B04组合的单果重中亲优势和超亲优势最为突出，分别达到42.47%和37.38%。     

Mode of action of abscisic acid in triggering ethylene biosynthesis and softening during ripening in mango fruit

The role of abscisic acid (ABA) in triggering ethylene biosynthesis and ripening of mango fruit was investigated by applying ABA [S-(+)-cis,trans-abscisic acid] and an inhibitor of its biosynthesis [nordihydroguaiaretic acid (NDGA)]. Application of 1 mM ABA accelerated ethylene biosynthesis through promoting the activities of ethylene biosynthesis enzymes (1-aminocyclopropane-1-carboxylic acid synthase, ACS; 1-aminocyclopropane-1-carboxylic acid oxidase, ACO) and accumulation of 1-aminocyclopropane-1-carboxylic acid (ACC), enhanced fruit softening and activity of endo-polygalacturonase and reduced pectin esterase activity in the pulp. The activities of ethylene biosynthesis and softening enzymes were significantly delayed and/or suppressed in the pulp of NDGA-treated fruit. The ABA-treated fruit had higher total sugars and sucrose as well as degradation of total organic acids, and citric and fumaric acids compared with NDGA treatment. These results suggest that ABA is involved in regulating mango fruit ripening and its effects are, at least in part, mediated by changes in ethylene production.     

Physiological response of ‘Larry Ann’ plums to cold storage and 1-MCP treatment

The aim of this work was to study the specific effects of low temperature and 1-MCP treatment on ethylene metabolism and oxidative behaviour in plums (Prunus × salicina cv. Larry Ann). Control fruit were stored at 20 °C or 0 °C and the 1-MCP (625 nL L^?1) treated fruit at 0 °C. Changes in the kinetics of ethylene production upon removal were related to changes in ACC metabolism (ACC and MACC levels), oxidative behaviour (H₂O₂ content) and enzymatic antioxidant potential (SOD, CAT and POX enzymes) during cold storage. Low temperature stress inhibited the synthesis of MACC, which appeared to be the basic process that regulated ACC and ethylene production at ambient temperature. Although 1-MCP treatment inhibited ethylene production and ACC accumulation in the cold, it did not inhibit the accumulation of MACC. Neither cold nor 1-MCP treatment induced oxidative stress. Nevertheless, the 1-MCP treatment significantly impaired the increase in POX activity observed during cold storage. Collectively these results showed the underlying role that ACC metabolism plays in the ripening behaviour of cold-stored plums, confirming previous results. The results also indicate that MACC and malonyl transferase activity are the key regulatory factors that control ripening and possibly some ethylene-related disorders such as chilling injury in cold-stored plums.     

甜瓜果皮颜色遗传分析及基因定位

为探究甜瓜皮色性状的遗传规律，定位目标性状关联的遗传位点，选用黄皮材料B432 和绿皮材料B168、B421 为亲本，构建6 世代遗传群体(P1、P2、F1、F2、BC1P1、BC1P2)，用于分析甜瓜皮色性状的遗传规律；同时，利用BSA-seq 和全基因组重测序技术，定位控制甜瓜皮色的基因。结果表明，甜瓜皮色由2对基因控制，其中绿色对白色具有显性上位性效应，白色对黄色为显性。同时，将皮色相关基因分别定位于4 号染色体和10 号染色体的0.02~5.7 Mb和0.08~9.5 Mb区间。试验初步定位了控制甜瓜皮色的基因，为后续进行基因精细定位提供依据，为开展甜瓜皮色的分子标记辅助选择奠定基础。