The role of the epidermis as a stiffening agent in Tulipa (Liliaceae) stems

We investigated the hypothesis that the epidermis is a tension-stressed "skin' whose contribution to stem stiffness depends on the turgor pressure exerted on it by an hydrostatically inflated inner "core' of tissues. This hypothesis was tested by relying on the intensities of bending stresses due to stem flexure, which must reach their maximum levels at the outer surface of epidermis such that damage to the surface of the stem should produce the most significant decrease in overall flexural stiffness. We discerned whether the principal tension supporting members at the stem surface (cellulosic microfibrils) were oriented parallel or normal to stem length by comparing the bending stiffness of stems before and after their surface cells first received three parallel longitudinal incisions followed by one helical incision, and by comparing the bending stiffness of stems for which the sequence of cuts was reversed. The same protocol was also applied to stems with various water potentials to determine the effect of hydrostatic pressure on stem stiffness contributed by the surface. Based on the behavior of 82 turgid Tulipa stems, parallel cuts reduced, on average, stem stiffness by 8%, whereas a subsequent helical incision further reduced stiffness by 42%. In contrast, an initial helical incision reduced stem stiffness by 50%, while three subsequent parallel cuts through the same stems did not significantly further reduce stiffness. These results suggested that the net orientation of cellulose microfibrils in the outer epidermal walls was parallel to stem length. This was confirmed by microscopic observations of cells with dichroic staining and polarized light. The responses to surgical damage were directly proportional to stem water potential. We thus conclude that the epidermis, probably in conjunction with a single layer of subepidermal collenchyma cells, acts as a tension-stiffening agent that can contribute as much as 50% to overall stem stiffness We present a simple mechanical model that can account for all our observations.     

郁金香衰老过程中几种保护酶活性的变化

两个郁金香品种在开花后随着可溶性蛋白含下降,细胞内超氧化物歧化酶、过氧化氢酶和过氧化物酶３种保护酶活性也随之下降,同时发生过氧化产物丙二醛的迅速累积;     

Changes in diffusible indole-3-acetic acid from various parts of tulip plant during rapid elongation of the flower stalk

In order to chemically identify the putative indole-3-acetic acid (IAA) and to confirm the native source of auxins account for rapid elongation of the floral stalk of tulip, we examined diffusible IAA from various parts of tulip plant during rapid elongation of the flower stalk. IAA was identified in the diffusates collected from the leaves, internodes, and floral organs with gas chromatography (GC)–mass spectrometry. The amount of diffusible IAA from different plant organs followed the order of that the internodes > flower organs > leaves during the period of rapid elongation of the floral stalk. The diffusible IAA from internodes reached its peak amount at different time than did diffusible IAA from the flower. The results obtained indicated that the top internode is probably the major source of auxins account for rapid elongation of the flower stalk.     

A vacuolar iron transporter in tulip, TgVit1, is responsible for blue coloration in petal cells through iron accumulation

Blue color in flowers is due mainly to anthocyanins, and a considerable part of blue coloration can be attributed to metal-complexed anthocyanins. However, the mechanism of metal ion transport into vacuoles and subsequent flower color development has yet to be fully explored. Previously, we studied the mechanism of blue color development specifically at the bottom of the inner perianth in purple tulip petals of Tulipa gesneriana cv. Murasakizuisho. We found that differences in iron content were associated with the development of blue- and purple-colored cells. Here, we identify a vacuolar iron transporter in T. gesneriana ( TgVit1 ), and characterize the localization and function of this transporter protein in tulip petals. The amino acid sequence of TgVit1 is 85% similar that of the Arabidopsis thaliana vacuolar iron transporter AtVIT1, and also showed similarity to the AtVIT1 homolog in yeast, Ca²⁺-sensitive cross-complementer 1 (CCC1). The gene TgVit1 was expressed exclusively in blue-colored epidermal cells, and protein levels increased with increasing mRNA expression and blue coloration. Transient expression experiments revealed that TgVit1 localizes to the vacuolar membrane, and is responsible for the development of the blue color in purple cells. Expression of TgVit1 in yeast rescued the growth defect of ccc1 mutant cells in the presence of high concentrations of FeSO₄. Our results indicate that TgVit1 plays an essential role in blue coloration as a vacuolar iron transporter in tulip petals. These results suggest a new role for involvement of a vacuolar iron transporter in blue flower color development.     

准噶尔盆地伊犁郁金香稳定碳同位素组成变化特征

选取准噶尔盆地古尔班通古特沙漠南缘9个样地中的伊犁郁金香（Tulipa iliensis）自然居群,通过测定各居群中伊犁郁金香的稳定碳同位素组成（δ^13C）、叶片和土壤中全氮、全磷、全钾以及水分含量,分析伊犁郁金香δ^13C值与上述因子之间的相互关系。结果表明：不同生境间伊犁郁金香δ^13C值差异不显著,水分利用方式可能受其基因型的控制而较为保守,同时在一定程度上也受生长环境中环境因子变化的影响,因而会有小的变幅：除了土壤中全钾含量之外,伊犁郁金香的叶片全氮、全磷、全钾、叶片水分含量以及土壤全氮、全磷和土壤水分含量在不同生境间均存在显著差异;其中伊犁郁金香叶片和土壤中的全氮含量与δ^13C值呈显著正相关：随着该植物叶片和土壤中水分含量的下降,伊犁郁金香δ^13C值显著增大,水分利用效率（WUE）明显提高。说明伊犁郁金香δ^13C值是植物的遗传学因子和环境因子共同作用的结果,其叶片及土壤中的氮素含量和有效水分含量是影响伊犁郁金香δ^13C值的关键因子。     

新疆野生种与栽培种郁金香的杂交种子无菌萌发和鳞茎形成

新疆野生种与栽培种郁金香的杂交F1种子于4℃低温下培养36-67d后萌发,随后于15℃下培养3-5个月形成鳞茎。‘小黑人’×柔毛郁金香、‘小黑人’×天山郁金香和‘小黑人’×伊犁郁金香F1种子的萌发率差异显著,分别为60．37％-63．89％、80．28％-81．25％和3．75％-4．45％。‘小黑人’×柔毛郁金香和‘小黑人’×天山郁金香F1的鳞茎分别在添加6-BA2．0mg·L^-1（单位下同）＋KT1．0＋TDZ0．1和6-BA5．0＋KT0．4的1／2MS培养基中增殖。‘小黑人’×天山郁金香F1种子萌发后有41．67％呈畸形,畸形的萌芽在添加6-BA2．0＋KT0．4的1／2MS培养基中可诱导出鳞茎并增殖。     

The effect of medium composition on ovary-slice culture and ovule culture in intraspecific Tulipa gesneriana crosses

The effect of several media components on the germination percentage of ovules in intraspecific T. gesneriana L. crosses was studied by using two embryo rescue techniques, viz. ovary-slice culture followed by ovule culture and direct ovule culture. The addition of 9% sucrose to medium for ovary-slice culture, started at 3 or at 5 weeks after pollination (WAP), significantly improved the germination percentage as compared to 5% sucrose. The germination percentage did not differ between both sucrose concentrations (3% and 5%) used in ovule culture started 4 weeks later with ovules excised from the ovary-slices (at 9 WAP). Similar germination percentages were obtained with media containing the full or half of the concentrations of micronutrients and macronutrients of the MS-medium during ovary-slice culture and ovule culture. For direct ovule culture, started at 4, at 6, and at 8 WAP, the germination percentages did not differ between ovules cultured on media with 3%, 6% or 9% sucrose. The addition of the cytokinin BAP (0.01 or 0.1 mg l^-1) had no effect on the germination percentage. The use of liquid-shaken culture resulted in germination percentages which were similar to those on agar-solidified media. Analysis of the carbohydrate concentration of the media revealed that, in both media for ovary-slice culture and for ovule culture, ultimately all sucrose is converted into glucose and fructose. The total concentration of carbohydrates decreased with 19%–48% in the media for ovary-slice culture, whereas the total concentration of carbohydrates did not decrease remarkably in media for ovule culture.     

Rapid stalk elongation in tulip (Tulipa gesneriana L. cv. Apeldoorn) and the combined action of cold-induced invertase and the water-channel protein γTIP

Many bulbous plants need a low-temperature treatment for flowering. Cold, for example, affects the elongation of the stalk, thereby influencing the quality of the cut flower. How the elongation of the stalk is promoted by cold and which physiological and biochemical mechanisms are involved have remained obscure. As invertase has been shown to be involved in the cold-induced elongation of the flower stalks of tulips (Lambrechts et al., 1994, Plant Physiol 104: 515–520), we further characterized this enzyme by cloning the cDNA and analysing its expression in various tissues of the tulip (Tulipa gesneriana L. cv. Apeldoorn) stalk. In addition, the role of sucrose synthase was investigated. Since turgor pressure is an important force driving cell elongation, the role of a water-channel protein (γTIP) was studied in relation to these two enzymes. The mRNA level of the invertase found was substantially up-regulated as a result of cold treatment. Analysis of the amino acid sequence of this invertase revealed the presence of a vacuolar targeting signal. Two different forms of sucrose synthase were found, the expression of one of them appeared to be restricted to the vascular tissue while the other form was present in the surrounding tissue. Both sucrose synthases were present in the stalk during the entire period of bulb storage and after planting, but their activities declined during stalk elongation. The expression of the γTIP gene was restricted mainly to the vascular tissue and its expression profile was identical to that of invertase. Simultaneous expression of invertase and γTIP possibly leads to an increase in osmotic potential and vacuolar water uptake, thus providing a driving force for stretching the stalk cells. Received: 12 January 1999 / Accepted: 24 March 1999     

Effects of gelling agents on in vitro regeneration and kanamycin efficiency as a selective agent in plant transformation procedures

Successful plant transformation requires effective regeneration and selection systems. The regeneration of tulip scape segments, gladiolus cormel slices, and tobacco leaf pieces were compared on media solidified with different gelling agents and with kanamycin at various concentrations. Increasing concentration of kanamycin generally resulted in full or partial inhibition of regeneration. However, regeneration was observed with one of the gelling agents, a κ-carrageenan, and 200 mg l⁻¹ kanamycin in the medium. With other gelling agents, 50% of this concentration was generally sufficient to totally inhibit regeneration. Therefore, the choice of the gelling agent is critical when establishing a plant transformation procedure. This revised version was published online in June 2006 with corrections to the Cover Date.     

Proton Efflux from the Outer Layer of the Peduncle of Tulip in Gravitropism and Circumnutation*

Epidermal plus hypodermal peels from tulip peduncles produced bands of acidity on agar containing bromocresol purple. Peels from horizontally oriented peduncles gave rise to an acidity band which corresponded to the lower side of the peduncle. The band began 3–6 cm beneath the flower and extended basipetally within the region of gravitropic bending. No corresponding band appeared in an agar layer laid on the cortical surface exposed by peeling. Peduncles growing in the normal vertical position showed circumnutations with a period in the range of 4 h. The peels from these stalks produced one or two bands more acid than the remaining part of the peel. Since the acidity band in horizontally positioned stalks corresponds to the zone of faster growth causing gravitropic bending, we infer that the band(s) produced by vertical stalks also correspond to zones of differential growth involved in circumnutation. On the basis of a previous finding that tulip leaves give rise to an oscillating acidity pattern, we infer that vertical stalks also show such a pattern. This inference fits the model proposing the involvement of an internal oscillator in circumnutation. However, the ratio of the circumnutation period to the gravitropic lag phase in tulip peduncles is such as predicted by the gravitropic-feedback model of circumnutation.