Down regulation of StGA3ox genes in potato results in altered GA content and affect plant and tuber growth characteristics

GA biosynthesis and catabolism has been shown to play an important role in regulating tuberization in potato. Active GAs are inactivated in the stolon tips shortly after induction to tuberization. Overexpression of a GA inactivation gene results in an earlier tuberization phenotype, while reducing expression of the same gene results in delayed tuberization. In addition, overexpression of genes involved in GA biosynthesis results in delayed tuberization, while decreased expression of those genes results in earlied tuberization. The final step in GA biosynthesis is catalysed by StGA3ox1 and StGA3ox2 activity, that convert inactive forms of GA into active GA₁ and GA₄. In this study we cloned StGA3ox2 gene in an RNAi construct and used this construct to transform potato plants. The StGA3ox2 silenced plants were smaller and had shorter internodes. In addition, we assayed the concentrations of various GAs in the transgenic plants and showed an altered GA content. No difference was observed on the time point of tuber initiation. However, the transgenic clones had increased number of tubers with the same yield, resulting in smaller average tuber weight. In addition, we cloned the promoter of StGA3ox2 to direct expression of the GUS reporter gene to visualize the sites of GA biosynthesis in the potato plant. Finally, we discuss how changes of several GA levels can have an impact on shoot, stolon and tuber development, as well as the possible mechanisms that mediate feed-forward and feed-back regulation loops in the GA biosynthetic pathway in potato.     

Jasmonic Acid induces tuberization of potato stolons cultured in vitro

The aim of the study was to assess the potential in vitro effects of jasmonic acid and kinetin on tuberization of potato (Solanum tuberosum). Of the two, the former was by far the stronger in vitro promoter of stolon tuberization. Number of tubers induced per stolon, tuberization rate, and final tuber weight were higher by factors of 2.8, 2.3, and 6.4, respectively. Bioassay sensitivity of jasmonic acid, measured in terms of the point at which the concentration for inducing tuberization was saturating, was more than 20 times greater than that of kinetin. Tuberization in both cases was associated with a decrease in rooting ability. Jasmonic acid also triggered a general state of induction throughout the stolon.     

Interaction between day length and phytohormones in the control of potato tuberization in the in vitro culture

We studied the interaction of the day length, cytokinins, and gibberellins in the control of tuberization in potato (Solanum tuberosum L, cv. Desire) plants and derived transgenic plants with the inserted PHYB gene from Arabidopsis encoding the synthesis of phytochrome B apoprotein and put under the control of the 35S CaMV promoter. Plantlets were cultured in vitro on hormone-free MS medium containing 5% sucrose and kinetin (1 mg/l) or/and GA (0.5 and 1.0 mg/l), at long day (LD, a 16-h photoperiod), short day (SD, a 10-h photoperiod), or continuous darkness conditions. The content of cytokinins (Ck, zeatin, and zeatin riboside) in various plant organs was determined by the immunoenzyme method, and GA activity was measured in bioassay with dwarf pea. Potato plant transformation with the PHYB gene enhanced substantially tuber initiation inhibition by LD. Kinetin addition to culture medium enhanced tuberization and reduced Ck content in aboveground shoots and Ck redistribution in the favor of underground organs. GA addition to the culture medium suppressed tuberization and induced Ck accumulation in aboveground organs. We concluded that Ck role in tuberization depends on their predominant localization in above- or underground potato organs. The involvement of Ck and GA in the competitive relations between growing tubers and shoots is considered.     

Relation between Nitrogen Nutrition, Cytokinin Activity and Tuberization in Solanum tuberosum

In water-culture experiments with potato plants (Solanum tuberosum L. cv. Ostara), the influence of tuberization initiated by a 7-day period of nitrogen withdrawal (discont. N) on the cytokinin activity in shoots, roots and exudate was studied. Plants with a constant supply of nitrogen (cont. N) were used as control. — Whereas no tuberization could be observed with cont. N, discont. N led to tuberization already 2 days after nitrogen withdrawal and all plants had been induced after another 4 days. In the roots of plants with discont. N, there was a temporary increase in cytokinin activity, whereas the activity decreased steadily with cont. N. In the exudate, cytokinin activity was greatly reduced during nitrogen withdrawal, whereas this activity in the exudate increased steadily with cont. N. — In the shoot with cont. N cytokinin activity decreased steadily, but with discont. N, after an initial decrease, the activity increased steeply. This increase is mainly or exclusively caused by a shift between the water-soluble and butanol-soluble fractions of the cytokinins in favour of the latter. The shift in cytokinin activity of the shoot is assumed to be in causal connection with an increased photosynthetic activity after the onset of tuber growth as ‘sink’ for assimilates.     

Cytokinin Activity in Stolons and Tubers of Solanum tuberosum during the Period of Tuberization

In water-culture experiments with potato plants (Solanum tuberosum L. cv. Ostara), changes in cytokinin activity in the stolon tips and newly formed tubers during tuberization were studied. Tuberization was induced by withdrawing nitrogen from the nutrient solution. — The cytokinin activity was low in the stolon tips prior to tuberization, but increased considerably in both stolon tips and young tubers during tuberization. At the same time qualitative changes in the cytokinin spectrum occurred. These qualitative changes are reversible if ‘regrowth’ of young tubers is brought about by a sudden high supply of nitrogen. — Despite the close correlation between tuberization and cytokinin activity, it is assumed that cytokinins are not directly responsible for the onset of tuberization, although they play an important role in tuber growth.     

丛枝菌根真菌对小麦生长的影响

为了促进经济作物小麦的生长,提高土壤氮磷循环与转化效率,选择两种优良丛枝菌根真菌(Arbuscular mycorrhizal fungi,AMF)——摩西球囊霉(GM,Glomus mosseae)、根内球囊霉(GI,Glomus intraradices),研究AMF在小麦整个营养生长阶段中对其生长以及对土壤中植物生长需求的大量元素——氮、磷的作用及影响。结果表明:人工施加菌剂可显著提高AMF对小麦的侵染率,施加GM菌剂时,小麦侵染率提高24.54%,同时,株高提高14.08%,小麦地上生物量提高24.05%。GM效果优于GI。施加菌剂后,小麦侵染率与土壤中水解性氮呈显著正相关;植物地上生物量与土壤中总氮,水解性氮呈显著负相关。表明AMF可活化土壤中的氮元素,同时促进作物生长,强化对土壤中氮元素的利用。     

A new approach for achievement of inulin accumulation in suspension cultures of Jerusalem artichoke (Helianthus tuberosus) using biotic elicitors

A promising protocol for achievement the accumulation rate of inulin compound in a suspension culture of Jerusalem artichoke (Helianthus tuberosus) was established. The effect of incorporated of cell cultures in combining with two type of biotic elicitors Aspergillus niger extract and Methyl-Jasmonate incorporation feeding medium on leaf cell growth patterns and production of inulin was investigated. The maximum value of cell growth parameters and highest content of inulinase activity (0.395 u/ml) were resulted from elicitation of augmented MS-medium with A. niger extract at the level of 0.2% in combination with Methyl-Jasmonate (150 μM) as compared with other concentrations after 2 weeks of cultivation. The chemical analyses of the different cell lines were spectro-photometerically performed. This study clearly indicates that combining of A. niger and Methyl-Jasmonate elicitors plays a critical role on inulin process and its accumulation in Jerusalem artichoke cell cultures.     

Physiology of Tuberization in Solanum tuberosum L: cis-Zeatin Riboside in the Potato Plant: Its Identification and Changes in Endogenous Levels as Influenced by Temperature and Photoperiod

Using high pressure liquid chromatography, the cucumber cotyledon bioassay, and mass spectrometry a cytokinin isolated from Solanum tuberosum L. cv. Katahdin plant tissues has been identified as cis-zeatin riboside. Zeatin riboside (ZR) levels in plants grown under inducing conditions (28 C day and 13 C night with a 10-hour photoperiod) were significantly higher than those in plants grown under noninducing conditions (30 C day and 28 C night with an 18-hour photoperiod). The highest level of ZR was noted in below-ground tissue after 4 days exposure to inducing conditions, with tuber initiation observed after 8 days. A companion study conducted to determine the effect of ZR on in vitro tuberization of noninduced rhizomes revealed that after 1 month in culture, controls exhibited 0% tuberization, while ZR treatments of 0.3 and 3.0 milligrams per liter showed 39 and 75% tuberization, respectively.     

Blue Light Inhibition of Tuberization in a Day-Neutral Potato

In tests on the effects of light quality on potato tuberization, continuous blue light was found to consistently inhibit tuberization of tissue-cultured plantlets of Solanum tuberosum ssp. tuberosum cv. ??Norland??. Other tested cultivars, including sports of ??Norland??, formed tubers under continuous blue light. Microarrays identified BL, GA7ox, and Nudix genes as exhibiting altered expression in response to blue light treatment. Quantitative RT-PCR (qRT-PCR) showed that GA7ox RNA increased in ??Norland?? but not in ??Sangre?? plantlets in blue light compared to darkness. RNA levels of genes identified in the literature as having roles in potato tuberization were also measured using qRT-PCR. Levels of GA20o1x, but not GA2ox, RNA increased in response to blue light in ??Norland?? plantlets. BEL5 RNA content was greater under blue light compared to darkness for both ??Norland?? and ??Sangre?? plants. Levels of FT were not significantly different in blue light compared to dark-treated ??Norland?? plants, but were low in blue light-treated compared to dark-treated ??Sangre?? plants. Addition of ancymidol to ??Norland?? plants exposed to blue light overcame blue light inhibition of tuberization. Ancymidol prevents the oxidation of ent-kaurene to ent-kaurenoic acid, thus inhibiting gibberellin biosynthesis. These data suggest that blue light may increase GA accumulation in ??Norland?? plants, as has been shown to occur in Arabidopsis plants. The novel effect of blue light in inhibiting tuberization of ??Norland?? plants suggests that this system could be a useful tool in further elucidating the mechanisms of day-neutral potato tuberization.     

测墒补灌对冬小麦氮素积累与转运及籽粒产量的影响

2007-2009年,在田间条件下,以冬小麦品种济麦22为材料,以0-140 cm土层平均土壤相对含水量为指标设计4个测墒补灌试验处理:W0(土壤相对含水量为播种期80%+拔节期65%+开花期65%)、W1(土壤相对含水量为播种期80%+拔节期70%+开花期70%)、W2(土壤相对含水量为播种期80%+拔节期80%+开花期80%)和W3(土壤相对含水量为播种期90%+拔节期80%+开花期80%),研究不同水分处理对冬小麦氮素积累与转运、籽粒产量、水分利用效率及土壤硝态氮含量的影响。结果表明:(1)成熟期小麦植株氮素积累量为W1处理最高,W3处理次之,W0和W2处理最低,W0和W2处理间无显著差异;氮素向籽粒的分配比例为W2处理显著低于W1处理,W0、W1、W3处理间无显著差异。开花期和成熟期营养器官氮素积累量、营养器官氮素向籽粒中的转移量、成熟期籽粒氮素积累量均为W1>W3>W2>W0,各处理间差异显著。(2)随着小麦生育进程的推进,0-200 cm土层土壤硝态氮含量先降低后回升再降低,在拔节期最低。成熟期W0和W1处理0-200 cm土层土壤硝态氮含量较低,W2和W3处理120-200 cm土层土壤硝态氮含量较高。(3)W0处理小麦氮素吸收效率、利用效率和氮肥偏生产力最低;随灌水量的增加,氮素利用效率呈先升高后降低趋势;W1处理小麦对氮素的吸收效率和利用效率较高,氮肥偏生产力最高。W0处理水分利用效率较高,但籽粒产量最低;灌水处理籽粒产量、灌溉水利用效率和灌溉效益两年度均随测墒补灌量的增加而显著降低。在本试验条件下,综合氮素利用、籽粒产量、灌溉水利用效率及土壤中硝态氮的淋溶,W1是高产节水的最佳灌溉处理,在2007-2008年和2008-2009年度补灌量分别为43.83 mm和13.77 mm。     

Effects of Nitrogen Nutrition on Nitrogen Partitioning between Chloroplasts and Mitochondria in Pea and Wheat

Nitrogen partitioning among proteins in chloroplasts and mitochondria was examined in pea (Pisum sativum L.) and wheat (Triticum aestivum L.) grown hydroponically with different nitrogen concentrations. In pea leaves, chloroplast nitrogen accounted for 75 to 80% of total leaf nitrogen. We routinely found that 8% of total ribulose-1,5-bisphosphate carboxylase/oxygenase adhered to thylakoids during preparation and could be removed with Triton X-100. With this precaution, the ratio of stroma nitrogen increased from 53 to 61% of total leaf nitrogen in response to the nitrogen supply, but thylakoid nitrogen remained almost constant around 20% of total. The changes in the activities of the stromal enzymes and electron transport in response to the nitrogen supply reflected the nitrogen partitioning into stroma and thylakoids. On the other hand, nitrogen partitioning into mitochondria was appreciably smaller than that in chloroplasts, and the ratio of nitrogen allocated to mitochondria decreased with increasing leaf-nitrogen content, ranging from 7 to 4% of total leaf nitrogen. The ratio of mitochondrial respiratory enzyme activities to leaf-nitrogen content also decreased with increasing leaf-nitrogen content. These differences in nitrogen partitioning between chloroplasts and mitochondria were reflected in differences in the rates of photosynthesis and dark respiration in wheat leaves measured with an open gas-exchange system. The response of photosynthesis to nitrogen supply was much greater than that of dark respiration, and the CO₂ compensation point decreased with increasing leaf-nitrogen content.     

Far-Red Reversal of Red Light Effect during Long-Night Induction of Potato (Solanum tuberosum L.) Tuberization

The hypothesis that phytochrome is involved in the regulation of potato (Solanum tuberosum L.) tuberization was tested. When 5 minutes of red light were given in the middle of the 16-hour dark period to which whole plants were exposed daily for 14 days before making cuttings, the percentage of tuberization on cuttings decreased. The effect of red light was significantly reversed by 2 minutes of far-red light given immediately after the red in each of two separate experiments. This supports the hypothesis that phytochrome is at least indirectly involved.     

The Relationship between Exogenous Growth Inhibitors and Endogenous Levels of Ethylene, and Tuberization of Dahlias

Treatments with abscisic acid (ABA), succinic acid-2,2-dimethylhydrazide (SADH), or 2 chlorethyl phosphonic acid (ethephon) promoted the tuberization of dahlia plants in long-days. This effect was smaller, however, than the effect short days have on tuberization. In contrast, gibberellic acid (GA) treatments inhibited tuberization. SADH and ethephon treatments of budless leaf-cuttings inhibited tuberization whereas ABA treatments slightly enhanced it. Evolution of endogenous ethylene reached a peak between the second and third week after the start of short-day treatments, and then decreased to the low level found in plants growing under long days. The peak in ethylene evolution occurred one week before the onset of tuberization.     

Photoperiodic and Hormonal Control of Tuberization in Potato Plants Transformed with the <Emphasis Type="Italic">PHYB</Emphasis> Gene from <Emphasis Type="Italic">Arabidopsis</Emphasis>

We studied photoperiodic and hormonal regulation of tuberization in wild-type potato (Solanum tuberosum L., cv, Desiree) plants and derivative transgenic plants harboring the PHYB gene from Arabidopsis, which encodes the phytochrome B apoprotein, under the control of the cauliflower mosaic virus 35S promoter. Plants were cultured on hormone-free Murashige and Skoog nutrient medium containing 5% sucrose or on the same medium supplemented with 1 mg/l kinetin under conditions of long day (LD, 16 h), short day (SD, 10 h), or SD with interrupted long night. We estimated cytokinins (zeatin and zeatin riboside) in underground and aboveground plant organs by the ELISA technique and GA activity in a bioassay with dwarf pea seedlings. Under LD conditions, transgenic plants produced substantially less tubers than wild-type plants. Kinetin addition to the culturing medium resulted in stimulation of tuberization under LD conditions, especially pronounced in the PHYB plants. The content of cytokinins and the activity of GA were much higher under LD conditions, especially in leaves. The total level of both phytohormones was higher in transformed as compared to wild-type plants. A relation of phytochrome-dependent tuberization to the hormonal status of underground and above-ground plant organs and possible reasons for kinetin stimulatory effect on this process are discussed.     

Efficient callus-mediated regeneration and <Emphasis Type="Italic">in vitro</Emphasis> root tuberization in <Emphasis Type="Italic">Trichosanthes kirilowii</Emphasis> Maxim., a medicinal plant

Trichosanthes kirilowii Maxim. is a climbing herb with considerable medicinal value. In this study, efficient protocols for callus-mediated regeneration and in vitro tuberization of this plant were developed. Sterilized stem and leaf tissues were cultured on Murashige and Skoog (MS) medium with plant growth regulators (PGRs), and additives that promoted callus induction and regeneration. Both stem and leaf tissues showed the best response (100%) for callus initiation on MS medium supplemented with 4.5-μM 2,4-dichlorophenoxyacetic acid (2,4-D). Efficient shoot organogenesis was obtained by exposing the callus tissue to 4.6-μM kinetin, 2.2-μM 6-benzylaminopurine, and 2.7-μM 1-naphthylacetic acid (NAA) along with 12.6-μM copper sulfate, which yielded a shoot regeneration rate of 85.5% and 28 shoots derived from each callus. In vitro shoots were best rooted on half-strength (1/2) MS medium with 2.7-μM NAA. Tuberous roots were efficiently induced on rooting medium with 5% (w/v) sucrose under short illumination conditions (8 h photoperiod). Rooted plantlets were successfully acclimatized in pots with a >?90% survival rate. This protocol provides an effective method for callus-mediated regeneration and in vitro root tuberization.     

Genetic stability of cultured hairy roots induced by Agrobacterium rhizogenes on tuber discs of potato cv. Bintje

Agrobacterium rhizogenes induced hairy roots on discs of the tuber tissue of the tetraploid potato cv. Bintje after infection. Early and late generated hairy roots (transformed roots) were excised directly from the tuber discs and analysed for genetic stability by chromosome counts and determination of nuclear DNA content with flow cytometry. Another part was cloned by subculturing on MS-medium without hormones and subsequently analysed. Twenty-one of the 27 primary hairy roots and all 16 subcultured hairy roots were tetraploid thus suggesting genetic stability of transformed roots. Shoot regeneration was observed on hairy roots and therefore, it is suggested that A. rhizogenes transformation can be a suitable system for genetically stable plant regeneration from transformed cells.     

A major QTL located on chromosome V associates with in vitro tuberization in a tetraploid potato population

The cultivated potato (Solanum tuberosum L.) is an autotetraploid species. The complexity of tetrasomic inheritance and the lack of pure lines increase the difficulty of genetic analysis of the inherited characteristics. Tuberization is the determinant step for economic yield of potato. To understand the complex genetic basis of tuberization of the cultivated potato, we developed linkage maps for a tetraploid population (F1) of 237 genotypes and mapped QTLs for the percent of in vitro tuberized plantlets (% IVT). The paternal map for E108 (well tuberized) covered 948 cM and included 12 linkage groups, all of which contained all four homologous chromosomes. The maternal map for E20 (nontuberized) covered 1,286 cM and included 14 linkage groups, 12 of which contained all four homologous chromosomes. All 12 chromosomes of potato were tagged using the SSR markers. A major QTL (MT05) with additive effect was detected on chromosome V of E108 which explained 16.23 % of the variation for % IVT, and two minor QTLs (mt05 and mt09) displaying simplex dominant effects were located on chromosome V and chromosome IX of E20 which explained 5.33 and 4.59 % of the variation for % IVT, respectively. Based on the additive model of MT05, the segregation ratio of the gametic genotypes (Q?: qq = 5:1) matched the ratio of the tuberized genotypes to the nontuberized genotypes in the population suggesting that the segregation of in vitro tuberization in this population is controlled by a major-effect gene or genes. The mapping results of three important candidate genes indicated that the QTL causal genes detected in our study are new. In this study, we developed the almost complete linkage maps of a tetraploid population, identified a major QTL on chromosome V affecting in vitro tuberization, suggested a major-effect gene with minor modifiers model controlling this trait and found that the QTLs identified here correspond to new tuberization genes. Our work provides new and useful information about the genetic basis for tuberization of this autotetraploid crop.     

亚洲玉米螟幼虫对氮、糖的吸收和利用

本文以不同含氮和含糖量的系列人工饲料为材料,研究了亚洲玉米螟Ostrinia fur nacalis(Guen6e)幼虫对氮和糖的吸收和利用。结果表明：幼虫对饲料中氮的吸收能力随虫龄增加而下降,对糖的吸收能力则随之而增强;5龄幼虫对糖的吸收能力随所食饲料中氮含量的增加而增强,而对氮的吸收则与饲料糖含量关系不大,5龄幼虫对氮和糖的消耗速率均随饲料含糖量增加而加快,但幼虫的氮利用率和虫体含氮量则随之下降;幼虫的氮消耗速率随饲料含氮量的增加而增加,而氮利用率则下降,5龄幼虫的氮利用率比3龄幼虫的氮利用率低,导致虫体含氮量的迅速下降。这些结果说明饲料中氮含量影响幼虫对糖的吸收,高龄幼虫对糖的需要量增加而对氮的需要则较稳定,饲料中糖比氮更重要。