Isolation and characterization of mRNAs differentially expressed during ripening of wild strawberry (Fragaria vesca L.) fruits

Wild strawberry (Fragaria vesca L.) is an attactive model system for studying ripening in non-climacteric fruit, because of its small diploid genome, its short reproductive cycle, and its capacity for transformation. We have isolated eight ripening-induced cDNAs from this species after differential screening of a cDNA library. The predicted polypeptides of seven of the clones exhibit similarity to database protein sequences, including acyl carrier protein, caffeoyl- CoA 3-O-methyltransferase, sesquiterpene cyclase, major latex protein, cystathionine -synthase, dehydrin and an auxin- induced gene. A ninth cDNA clone that was constitutively expressed is predicted to encode a metallothionein-like protein. None of these proteins appear to be directly related to events generally associated with ripening such as cell wall metabolism or the accumulation of sugars and pigments, rather, their putative functions are indicative of the wide range of processes upregulated during fruit ripening.     

Methyl Jasmonate Reduces Water Stress in Strawberry

The effect of methyl jasmonate (MJ) on changes of oxygen-scavenging enzyme activities and membrane lipid composition was studied in strawberry leaves under water stress. Under water stress, MJ treatment reduced the increase of peroxidase (EC 1.11.1.7; POD) activity, maintained higher catalase (EC 1.11.1.6; CAT) and superoxide dismutase (EC 1.15.1.1; SOD) activities, and ascorbic acid content. In addition, MJ treatment reduced transpiration and membrane-lipid peroxidation as expressed by malondialdehyde (MDA) content, lessened the reduction of membrane lipids, glycolipids [monogalactosyl diglyceride (MGDG), digalactosyl diglyceride (DGDG)], and phospholipids [phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), and phosphatidylinositol (PI)]. In water-deficit conditions, MJ treatment also alleviated the decline in the degree of fatty acid unsaturation and the ratio of linolenic (18:3) to linoleic acid (18:2). These results indicate that MJ treatment appears to alter the metabolism of strawberry plants rendering the tissue better able to withstand water stress. Received June 16, 1999; accepted October 1, 1999     

四种蜜源植物花蜜腺的比较解剖学研究

草莓、漆树、芥菜和薄荷四种植物花蜜腺形态结构各异。草莓花蜜腺着生在花管内壁上属花被蜜腺。芥菜花蜜腺４枚,包括一对侧蜜腺和一对中蜜腺。漆树和薄荷的花蜜腺为花盘蜜腺,位于子房基部。四种花蜜腺都属结构蜜腺,其中仅芥菜的侧蜜腺组织中具维管束。它们在发育早期均无特殊的原始细胞,都由花器官表层细胞反分化形成原基,再发育成蜜腺。四种花蜜腺发育过程中细胞原生质体的液泡都表现出了规律性的消长变化。漆树和薄荷的花蜜腺具典型的淀粉动态变化,草莓的花蜜腺则属非淀粉型蜜腺;芥菜花蜜腺介于这两种类型之间。草莓和薄荷的花蜜由表皮细胞直接渗出。芥菜则气孔和表皮泌出兼备而漆树通过气孔泌蜜。     

资源交互斑块性生境中两种不同分枝型匍匐茎植物的克隆内分工

以青藏高原和四川盆地过渡带两种不同分枝型匍匐茎植物野草莓 (Fragaria vesca)和过路黄 (Lysimachia christinae)为对象 ,研究它们在高光照低养分斑块和低光照高养分斑块组成的资源交互斑块性生境中的克隆内分工。结果显示 ,与资源的空间同质性处理 (I)和 (II)相比 ,资源的空间异质性处理 (III)和 (IV)中野草莓和过落黄的近端、远端和整个克隆片段的生物量和分株数均获得显著增加。生长在低光高养条件下的远端分株 ,若与高光低养的近端分株相连 ,相比连接到低光高养的近端分株 ,它们分配更多的生物量到地下部分 ;生长在高光低养条件下的远端分株 ,若与低光高养的近端分株相连 ,相比连接到高光低养的近端分株 ,它们分配更多的生物量到地上部分 ;生长在高光低养条件下的近端分株 ,若与低光高养的远端分株相连 ,相比连接到高光低养的远端分株 ,它们分配更多的生物量到地上部分。实验结果表明 ,资源交互斑块性生境中野草莓和过路黄均发生了克隆内分工。通过克隆内分工 ,克隆植物能有效的利用异质性分布的资源 ,缓解资源交互斑块性分布对克隆植物生长的不利影响     

Direct adventitious shoot formation on seedling radicles in seed cultures of strawberry

Mature seeds of strawberry (Fragaria x ananassa) were placed on Murashige and Skoog medium supplemented with 2.22 μM 6-benzyladenine. After four weeks of culture, and without an intervening callus phase, approximately 36% of the resulting seedling radicles had formed numerous adventitious buds near their tips. A few buds on each radicle developed into shoots, while others formed disorganized calli. Consequently, the seedlings exhibited shoot apices at both ends of the axis of polarity. Our overall results suggest that a considerable level of plasticity in organ determination occurs even in higher plants, and that exogenous growth regulators can cause a root primordium in the radicle to be converted to a shoot primordium.     

The influence of photoperiodic growth condition on isolation of RNA from strawberry (Fragaria × ananassa duch.) tissue

Purification of high-quality RNA from different strawberry tissues is often affected by the presence of high levels of contamination by polysaccharides and phenolic compounds. With the protocol detailed here we describe for the first time total RNA purification from petiole tissue. Treating the plants used as source of material with short-day light regime prior the extraction we are able to obtain RNA suitable for further applications such as in vitro translation, RT-PCR, and RNA blot analysis. The yield of total RNA extraction is significantly enhanced when tissue from plants grown under short-day photoperiodic condition is used compared with that taken from plants grown under long day photoperiod.     

Transposon tagging in diploid strawberry

Fragaria vesca was transformed with a transposon tagging construct harbouring amino terminally deleted maize transposase and EGFP (Ac element), NPTII, CaMV 35S promoter (P35S) driving transposase and mannopine synthase promoter (Pmas) driving EGFP (Ds element). Of 180 primary transgenics, 48 were potential launch pads, 72 were multiple insertions or chimaeras, and 60 exhibited somatic transposition. T(1) progeny of 32 putative launch pads were screened by multiplex PCR for transposition. Evidence of germ-line transposition occurred in 13 putative launch pads; however, the transposition frequency was too low in three for efficient recovery of transposants. The transposition frequency in the remaining launch pads ranged from 16% to 40%. After self-pollination of the T(0) launch pads, putative transposants in the T(1) generation were identified by multiplex PCR. Sequencing of hiTAIL-PCR products derived from nested primers within the Ds end sequences (either P35S at the left border or the inverted repeat at the right border) of T(1) plants revealed transposition of the Ds element to distant sites in the strawberry genome. From more than 2400 T(1) plants screened, 103 unique transposants have been identified, among which 17 were somatic transpositions observed in the T(0) generation. Ds insertion sites were dispersed among various gene elements [exons (15%), introns (23%), promoters (30%), 3' UTRs (17%) as well as intergenically (15%)]. Three-primer (one on either side of the Ds insertion and one within the Ds T-DNA) PCR could be used to identify homozygous T(2) transposon-tagged plants. The mutant collection has been catalogued in an on-line database.     

Localization of strawberry (Fragaria x ananassa) and Methylobacterium extorquens genes of strawberry flavor biosynthesis in strawberry tissue by in situ hybridization

Strawberry flavor is one of the most popular fruit flavors worldwide, with numerous applications in the food industry. In addition, the biosynthetic origin of the most important strawberry flavor components, such as 2,5-dimethyl-4-hydroxy-2H-furan-3-one (DMHF), is a challenging research area. DMHF's precursor, 2-hydroxy-propanal (or lactaldehyde), is biosynthesized by the endophytic bacterium Methylobacterium extorquens (M. extorquens). In particular, the alcohol dehydrogenase (ADH) enzymes of M. extorquens are involved in the biogenesis of DMHF precursors since they have the capacity to oxidize the strawberry-derived 1,2-propanediol to lactaldehyde. In this study, the expression of the endophytic ADH and the plant DMHF biosynthesis genes was examined in the tissues of raw and ripe strawberry receptacles by in situ hybridization. The presence of endophytic bacteria was studied in the same tissues by probes targeting bacterial 16S ribosomal ribonucleic acid. Hybridization signals of probes specific for endophytic ADH and plant DMHF biosynthesis genes, as well as bacteria-specific probes, were detected in the same locations. The probes were localized near the plasma membranes or intercellular spaces of cortical and vascular tissues of the receptacle, and intracellularly in the tissues of achenes. By localizing the expression of the endophytic methanol ADH and plant DMHF biosynthesis genes to the same tissues, we have reinforced our original hypothesis that an intimate symbiotic relationship between strawberry and endophytic cells exists and leads to the biosynthesis of DMHF.