Cloning and Sequencing the Full-length cDNA of Annexin from Strawberry Fruit

草莓果实膜联蛋白基因（annfaf）全长cDNA克隆及序列分析 王关林¹杨怀义^1,2*夏然¹ 方宏筠¹景士西³     

Respiration and plasma membrane ATPase in strawberry stolons

The biochemical, physiological and anatomical properties of strawberry (Fragaria ananassa Duch.) cv. 'Cambridge Favourite' stolons were studied during growth. ATPase activity was measured, in microsomal and plasma membrane fractions, along with chlorophyll determination, in-situ photosynthesis measurements and scanning electron microscopy (SEM) and X-ray microanalysis of stolon cross-sections. Potassium-stimulated ATPase activity and proton-pumping, both together indicating the presence of plasma membrane ATPase, was greatest in the stolon tip, the tissue with the fastest growth and respiratory activity. The enzyme activity and respiration gradient from the tip of the stolon to the base was concomitant with xylem development which was more differentiated in the base than in the tip. These cross-sections also showed 30% greater amounts of calcium and potassium of the cryo-preserved basal part relative to the stolon tip. This gradient existed independent of the presence of daughter plants. A hypothesis is presented which suggests that for the long-distance longitudinal transport of nutrients this gradient between stolon tip and base is likely to be involved in stolon growth.     

Effect of ammonium or nitrate nutrition on net photosynthesis,growth, and activity of the enzymes nitrate reductase and glutamine synthetase in blueberry,raspberry and strawberry

Blueberry, raspberry and strawberry may have evolved strategies for survival due to the different soil conditions available in their natural environment. Since this might be reflected in their response to rhizosphere pH and N form supplied, investigations were carried out in order to compare effects of nitrate and ammonium nutrition (the latter at two different pH regimes) on growth, CO2 gas exchange, and on the activity of key enzymes of the nitrogen metabolism of these plant species. Highbush blueberry (Vaccinium corymbosum L. cv. 13–16–A), raspberry (Rubus idaeus L. cv. Zeva II) and strawberry (Fragaria × ananassa Duch. cv. Senga Sengana) were grown in 10 L black polyethylene pots in quartz sand with and without 1% CaCO3 (w: v), respectively. Nutrient solutions supplied contained nitrate (6 mM) or ammonium (6 mM) as the sole nitrogen source. Compared with strawberries fed with nitrate nitrogen, supply of ammonium nitrogen caused a decrease in net photosynthesis and dry matter production when plants were grown in quartz sand without added CaCO3. In contrast, net photosynthesis and dry matter production increased in blueberries fed with ammonium nitrogen, while dry matter production of raspberries was not affected by the N form supplied. In quartz sand with CaCO3, ammonium nutrition caused less deleterious effects on strawberries, and net photosynthesis in raspberries increased as compared to plants grown in quartz sand without CaCO3 addition. Activity of nitrate reductase (NR) was low in blueberries and could only be detected in the roots of plants supplied with nitrate nitrogen. In contrast, NR activity was high in leaves, but low in roots of raspberry and strawberry plants. Ammonium nutrition caused a decrease in NR level in leaves. Activity of glutamine synthetase (GS) was high in leaves but lower in roots of blueberry, raspberry and strawberry plants. The GS level was not significantly affected by the nitrogen source supplied. The effects of nitrate or ammonium nitrogen on net photosynthesis, growth, and activity of enzymes in blueberry, raspberry and strawberry cultivars appear to reflect their different adaptability to soil pH and N form due to the conditions of their natural environment. This revised version was published online in July 2006 with corrections to the Cover Date.     

EST‐derived polymorphic microsatellites from cultivated strawberry (Fragaria×ananassa) are useful for diversity studies and varietal identification among Fragaria species

Microsatellite or simple sequence repeat markers derived from expressed sequence tags (ESTs) provide genetic markers within potentially functional genes, which could be very useful for breeding programs. To date, the development of microsatellite markers in the genus Fragaria has focused mainly on Fragaria vesca. However, most of the interests of breeding programs relate to specific characteristics of cultivated strawberry. Here, we describe a set of 10 EST‐derived microsatellites from Fragaria × ananassa. These markers showed high levels of polymorphism within strawberry cultivars and among different Fragaria species, indicating their potential for genetic studies not only on strawberry but also in other species within the genus.     

Ultrastructural changes in strawberry leaves infested by two-spotted spider mites

Ultrastructural changes in strawberry leaves after different periods of feeding by Tetranychus urticae were studied. Electron micrographs indicate that the first detectable changes in cells directly punctured by the mites usually occurred after 3 days of feeding and were confined to the chloroplasts. These organelles show instability of the lamellae and the thylakoid membrane system as well as the envelope. Longer times of mite feeding (6–9 days) caused severe alterations, not only within chloroplast. In heavily injured tissues, misshapen cells contain homogeneous protoplasts in which only remains of necrotic chloroplasts were visible. Mesophyll cells adjacent to directly punctured tissues also exhibited subcellular alterations. Possible mechanisms of plant-tissue responses to mite feeding are discussed.

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Zusammenfassung Ultrastrukturelle Veränderungen wurden in Erdbeerblättern nach verschiedenen Perioden Saugtätigkeit von Tetranychus urticae Koch gesucht. Elektronenmikroskopische Aufnahmen zeigen, dass die ersten sichtbaren Veränderungen in direkt von Milben angestochenen Zellen meist nach 3 Tagen Saugen auftraten und auf die Chloroplasten beschränkt waren. Diese Organellen zeigen eine Instabilität der Lamellen, des Thylakoidmem-bransystems sowie der Umhüllung. Ein längere Zeit dauerndes Saugen der Milben (6–9 Tage) verursachte schwere Veränderungen und zwar nicht bloss in den Chloroplasten. In schwer geschädigten Geweben enthalten deformierte Zellen homogene Protoplasten, in denen nur Reste nekrotischer Chloroplasten sichtbar waren.Mesophyllzellen in der Nachbarschaft von direkt angestochenem Gewebe zeigte ebenfalls subzelluläre Veränderungen. Mögliche Mechanismen der Reaktion des Pflanzengewebes auf Milbensaugen werden diskutiert.

     

Microsatellite markers for Fragaria from ‘Strawberry Festival’ expressed sequence tags

We present 37 microsatellite primer pairs developed from a cDNA library of Fragaria ^xananassa Duch. cv. Strawberry Festival. Polymorphism was high and the number of presumptive alleles of 13 expressed sequence tag–simple sequence repeats (EST–SSRs) in 70 strawberry cultivars ranged from five to 32 per primer pairs, averaging 16.1. Cross‐species amplification was also high and ranged from 89% in Fragaria vesca L. to 100% in the progenitor species of octoploid strawberry, Fragaria chiloensis (L.) Duch. and Fragaria virginiana Duch.     

Induced expression of the Fragaria × ananassa Rapid alkalinization factor-33-like gene decreases anthracnose ontogenic resistance of unripe strawberry fruit stages

Rapid alkalinization factor (RALF) genes encode for ubiquitous small peptides that stimulate apoplastic alkalinization through interaction with malectin-like receptor kinase. RALF peptides may act as negative regulators of plant immune response, inhibiting the formation of the signal receptor complex for immune activation. Recently RALF homologues were identified in different fungal pathogen genomes contributing to host infection ability. Here, FaRALF-33-like gene expression was evaluated in strawberry fruits inoculated with Colletotrichum acutatum, Botrytis cinerea, or Penicillium expansum after 24 and 48 h post-infection. To investigate the role of FaRALF-33-like in strawberry susceptibility, transient transformation was used to overexpress it in white unripe fruits and silence it in red ripe fruits. Agroinfiltrated fruits were inoculated with C. acutatum and expression, and histological analysis of infection were performed. Silencing of FaRALF-33-like expression in C. acutatum-inoculated red fruits led to a delay in fruit colonization by the fungal pathogen, and infected tissues showed less penetrated infective hyphae than in wild-type fruits. In contrast, C. acutatum-inoculated white unripe fruits overexpressing the FaRALF-33-like gene decreased the ontogenic resistance of these fruits, leading to the appearance of disease symptoms and penetrated subcuticular hyphae, normally absent in white unripe fruits. The different response of transfected strawberry fruits to C. acutatum supports the hypothesis that the FaRALF-33-like gene plays an important role in the susceptibility of fruits to the fungal pathogen C. acutatum.     

The control of red colour by a family of MYB transcription factors in octoploid strawberry (Fragaria × ananassa) fruits

Octoploid strawberry (Fragaria × ananassa Duch.) is a model plant for research and one of the most important non‐climacteric fruit crops throughout the world. The associations between regulatory networks and metabolite composition were explored for one of the most critical agricultural properties in octoploid strawberry, fruit colour. Differences in the levels of flavonoids are due to the differences in the expression of structural and regulatory genes involved in flavonoid biosynthesis. The molecular mechanisms underlying differences in fruit colour were compared between red and white octoploid strawberry varieties. FaMYB genes had combinatorial effects in determining the red colour of fruit through the regulation of flavonoid biosynthesis in response to the increase in endogenous ABA at the final stage of fruit development. Analysis of alleles of FaMYB10 and FaMYB1 in red and white strawberry varieties led to the discovery of a white‐specific variant allele of FaMYB10, FaMYB10‐2. Its coding sequence possessed an ACTTATAC insertion in the genomic region encoding the C‐terminus of the protein. This insertion introduced a predicted premature termination codon, which suggested the loss of intact FaMYB10 protein playing a critical role in the loss of red colour in white octoploid strawberry.     

利用黄毛草莓创制蜜桃香味草莓新株系

为创制具有蜜桃香味的草莓新株系,用栽培品种‘红颊’（‘Benihoppe’,2n=8x）与黄毛草莓（Fragaria nilgrrensis,2n=2x）为亲本进行杂交,得到种间杂种五倍体（2n=5x）,再经秋水仙素诱导后获得十倍体（2n=10x）,收获该十倍体的种子进行实生选种,选育出4个具有蜜桃香味的草莓新株系。结果表明,‘红颊’×黄毛草莓的杂交组合结实率和出苗率分别为39.58%和28.11%,适宜草莓十倍体诱导的秋水仙素浓度在200 mg/L左右,处理时间30~40 d,4个草莓新株系均具有蜜桃香味,抗氧化物质含量显著提高,但果肉质地较松软,平均单果重和株产稍低。这为野生草莓资源的利用和种质创新提供了科学参考。