Flowering and gibberellins in a mutant red clover (Trifolium pratense L.)

Relationships between gibberellins and floral initiation were investigated in a conditional non-flowering mutant of red clover, Trifolium pratense. Untreated mutant plants will not flower under long-days, but will do so when certain GAs are applied. Gibberellins, A₃, A₁, A₇, and A₅ all resulted in both stem elongation and flowering whilst GA₄ produced the elongation only. Applications of GA₂₀, GA₈ and GA₁₃ under long-days had no detectable effect. Thus, by combining the use of the mutant with the application of different GAs, the correlation between the processes of stem elongation and floral initiation, which is normally strongly expressed in this species, was broken. Endogenous gibberellins shown to be present in normal plants were also found in the mutant genotype. Gibberellins alone were not sufficient to initiate floral development in the mutant, there being an essential element of interaction with long-days. These results are discussed in relation to the nature of the lesion in the mutant and the signal provided by the applied gibberellin.     

Genotype-dependent whole plant regeneration from protoplasts of red clover (Trifolium pratense L.)

Protoplasts are useful for subcellular studies, in vitro selection, somatic hybridization and transformation. Whole plant regeneration from protoplasts is a prerequisite to producing altered crop plants using these methods. Whole plant regeneration was achieved from leaf- and suspension culture-derived protoplasts of T. pratense. Regeneration was most dependent upon identifying genotypes with genetic capacity to regenerate. Additional factors that were used to select genotypes, but which proved to be less important, were a high rate of cell growth in culture and a high plating efficiency of protoplasts. One genotype was identified which had a regeneration response equivalent to that of T. rubens and which regenerated from both leaf- and suspension culture-derived protoplasts.Research supported by USDA/CRGO Grant No. 81 CRCR-1-0613     

施用有机肥对高砷红壤中小白菜砷吸收的影响

通过盆栽试验,研究了施用猪粪和鸡粪条件下红壤中的砷对小白菜生长和吸收的影响及土壤有效态砷含量的变化.结果表明：向高砷红壤中施用猪粪和鸡粪两种有机肥均使小白菜的生物量有不同程度的增加,其中,施用猪粪处理的小白菜生物量显著高于对照处理(P<0.05);猪粪和鸡粪两种有机肥的施用可导致土壤有效砷含量明显提高,施用猪粪后土壤有效砷含量增幅达394.9%～1033.6%,而施用鸡粪的土壤有效砷含量增幅为30.4%～94.1%; 施用有机肥明显促进了小白菜对砷的吸收,其中猪粪处理下小白菜的砷吸收量比对照增加20.7%～53.9%. 根据本研究结果,对砷含量较高的高风险农田,施用猪粪、鸡粪等有机肥可能会在一定程度上提高土壤有效砷含量和作物对砷的吸收量,使农产品质量和环境风险增加.     

Effects of seed phosphorus concentration on the emergence and growth of subterranean clover (Trifolium subterraneum)

Subterranean clover seed (Trifolium subterraneum cv. Dalkeith) with phosphorus concentrations of 0.75% (high P seed) and 0.48% (low P seed) and of uniform size (2.0–2.4 mm diameter) was used to measure the effect of seed P concentrations on seedling emergence and growth.Seedling emergence numbers were 35% greater for the high P seed, and this effect was independent of external P supply. High P seed also emerged more quickly than low P seed.Leaf emergence was faster and shoot dry weight was greater for seedlings grown from high compared with low P seed, but only when external P supply was deficient for plant growth. Phosphorus concentrations in the shoots of two-week old seedlings were 32–51% higher for high P seed, although by four weeks plants grown from high and low P seed had similar concentrations of P in their shoots. We suggest that establishing pastures using high P seed would improve both early and late season pasture production.     

Artificial synthesis of interspecific chimeras between tuber mustard (Brassica juncea) and cabbage (Brassica oleracea) and cytological analysis

Interspecific chimeras between tuber mustard and red cabbage were obtained by in vitro graft-culture method. Before grafting, 6-day-old seedlings of tuber mustard and red cabbage were vertically half-cut and treated with different concentrations of 6-BA and NAA for 1 min, then, they were symmetrically fit together. As a result, sectorial chimeras were initially produced from the united shoot tips. The maximum frequency of chimeral bud formation reached 6.33% when the vertical sections of tuber mustard and cabbage were treated with 2 mg/l 6-BA and 1 mg/l NAA. When sectorial chimeras were propagated on MS medium containing 1 mg/l 6-BA, periclinal and mericlinal chimeras gradually developed. Chimeral shoots were rooted on half-strength MS medium containing 0.1 mg/l NAA. The rooted chimeras were acclimatized and transferred to the field for cytological and morphological analysis. The results showed that stomata density in the chimeras was significantly higher than that of their parents, while chloroplast size, starch grain size and number were intermediate between the two parents. The chimeras were further analyzed by flow cytometry, and the results indicated that they contained both sets of parental chromosomes. Moreover, chimeral plants possessed valuable characters from the two parents.     

The influence of long-term tillage systems on symbiotic N2 fixation of pea (Pisum sativum L.) and red clover (Trifolium pratense L.)

Pea as a grain legume and red clover as a forage legume in the seeding year were cultivated in two long-term differentiated tillage systems on a loess soil in Germany. A continuous conventional tillage system (plow; CT) and a continuous minimum tillage system (rotary harrow; MT) were established in 1970. With pea and red clover dry matter accumulation and N parameters (N accumulation, Ndfa, N-harvest-index, N balance) were investigated in 1998 and 1999. Differences in the N₂ fixation of pea due to the tillage system could clearly be shown whereas grain yields and total N accumulation were equal in both tillage systems and years. In both years a significantly (P < 0.05) higher Ndfa in the MT system was found at least in the final harvest (maturity of pea): 1998/1999, 0.42/0.54 in CT, 0.62/0.75 in MT. The differences in N₂ fixation of pea may be explained by the delayed soil N supply in MT at the beginning of the vegetative period. Simplified N balances of pea were -18 and –25 kg N ha^–1 in CT and –5 and +1 kg N ha^–1 in MT for 1998 and 1999, respectively. Red clover showed no significant differences in the DM and N accumulation between both tillage systems but a year dependent effect caused by different stubble and root yields between the years was apparent. With red clover slightly, but also significantly (P < 0.05) increased Ndfa values were found in the MT system compared to the CT system with 0.55/0.62 in CT (1998/1999) and 0.64/0.71 in MT. However, the difference in Ndfa between the tillage systems (9 percentage points) was much smaller with red clover than with pea (20 and 21 percentage points in 1998 and 1999, respectively). Soil N uptake of red clover using the longer growing season reflected the more adjusted N supply in both long-term differentiated tillage systems, whereas pea in using only a short-term vegetative period reacted stronger to the lower N mineralization in the MT system in springtime.     

Alternaria incidence in some alloplasmic lines of Indian mustard (Brassica juncea (L.) Coss.)

Summary The cytoplasmic substitution lines of Brassica juncea (L.) Coss were evaluated for their field resistance to Alternaria blight (Alternaria brassicae). The euplasmic B. juncea cv. RLM 198 had a mesothetic reaction while alloplasmic B. juncea lines with cytoplasms of B. campestris, B. chinensis, and B. japonica were highly susceptible. B. nigra cytoplasm did not have any effect on the disease reaction of the B. juncea genome. However, the alloplasmic lines with the cytoplasm of B. napus and B. carinata revealed a comparatively higher degree of resistance. The study underlined the utility of cytoplasmic manipulations in modifying the phenotypic expression of nuclear genes.     

轮作及绿肥不同利用方式对作物产量和土壤肥力的影响

通过4年田间定位试验比较了3种轮作及相应绿肥不同利用方式对作物产量和土壤性质的影响.轮作方式包括夏休闲-冬小麦（对照）、豆类绿肥-冬小麦和豆类绿肥-春玉米-冬小麦.豆类绿肥-冬小麦包括3种绿肥利用方式：提前覆盖、提前翻压和播前翻压;豆类绿肥-春玉米-冬小麦也包括3种绿肥利用方式：豆类茎秆覆盖、茎秆翻压和茎秆移出田间.结果表明：对于豆类绿肥-冬小麦轮作,绿肥消耗了更多小麦播前土壤水,使小麦产量不稳定;麦收后0～200 cm土层硝态氮储量显著高于另外两种轮作,有更高的淋失风险;该轮作方式下提前覆盖处理0～20 cm土层土壤有机碳(SOC)含量和有机碳储量(SSOC)最高.对于豆类绿肥-春玉米-冬小麦轮作,小麦播前土壤储水量显著高于豆类绿肥-冬小麦,小麦产量更稳定;麦收后0～200 cm土层硝态氮储量显著低于豆类绿肥-冬小麦轮作,淋失风险较低;该轮作方式下茎杆覆盖处理0～20 cm土层土壤SOC含量显著高于茎杆移出处理,且SSOC相对于试验初始也有所增加.可见,豆类绿肥-春玉米-冬小麦轮作体系中豆类收获籽粒后茎杆地表覆盖方式,在提高小麦播前土壤储水量、稳定产量、培肥土壤和降低0～200 cm土层土壤硝态氮残留量上表现较好,是具有类似气候地区的合理种植制度.
     

豆科绿肥及施氮量对旱地麦田土壤主要肥力性状的影响

通过2a田间定位试验,研究渭北旱塬地区夏闲期插播并翻压不同豆科绿肥(长武怀豆、大豆和绿豆)以及小麦生长季不同施氮量(0,108,135,162 kg/hm2)对麦田土壤肥力性状的影响,以期为提高旱地土壤质量提供理论依据.试验结果表明:(1)种植豆科绿肥能显著提高土壤有机质、活性有机质和全氮含量,增加土壤碳库管理指数(CPMI),对土壤速效钾含量没有显著影响;(2)绿豆还田量高于长武怀豆和大豆,然而土壤培肥效果逊于长武怀豆和大豆;(3)夏闲期种植绿肥明显消耗了土壤水分,导致绿肥翻压前、小麦播前直至收获后,0-200 cm土壤贮水量显著低于休闲处理,但耗水量与休闲没有明显差异,由于小麦产量显著增加,因此豆科绿肥显著提高了水分生产效率;(4)与不施氮相比,小麦生长季施用氮肥能显著增加土壤水分生产效率,却对土壤各肥力性状的影响均不显著.夏闲期种植并翻压豆科绿肥是旱地培肥土壤、提高水分生产效率的有效途径.     

Effect of mustard green manure and dried plant residue on chickpea wilt (Fusarium oxysporum f.sp. ciceris)

Pot experiments were carried out in the green house at Amhara Regional Agriculture Research Institute (ARARI) Bahirdar, Ethiopia, to evaluate the potential of Brassica carinata cultivars, namely Holleta-l, S-67 and Yellow Dodola in 2007 and 2008. The effect of B. carinata (Ethiopian mustard) cultivars Holleta-1, S-67 and Yellow Dodola as green manure and Holleta-1 as dried plant residue on chickpea fusarium wilt (Fusarium oxysporum f.sp. ciceris) was studied. Six rates of green manure and dried plant residue (0, 20, 40, 60, 80 and 100 g) each per kg of pathogen-infested soil were used in the experiments. Infested soil without B. carinata cultivars amendment as a control and susceptible check variety JG-62 without amendment was used in the experiments. In the experiments, the treatments were arranged in randomised complete block design in three replications and repeated twice. Data on seedling emergence, wilt incidence, fresh weight and dry weight were collected. The amendments of infested soil with B. carinata cultivars green manure and dried plant residue reduced the incidence of chickpea fusarium wilt. The incorporation of the green manure Holleta-1, S-67 and Yellow Dodola at 20–100 g/kg of infested soil was effective in reducing wilt incidences on chickpea. However, the incorporation of Yellow Dodola at 80 and 100 g green manure per kg of infested soil were the best combination in reducing significantly wilt incidence. The application of the dried plant residue at 20–100 g/kg of infested soil was effective in reducing wilt incidences on chickpea. However when applied dried plant residue at 60, 80 and 100 g green manure per kg of infested soil were better in reducing wilt incidence as compared to 20 and 40 g/kg of infested soil. The three cultivars green manure incorporated at different level of doses affected the influence of fusarium wilt on the fresh and dry weight respectively. The use of Holleta-1 green manure at 20–100 g/kg of infested soil significantly reduced disease incidence in the range of 20.0–33.3%. Green manure amendment S-67 significantly reduced disease incidence in the range of 20.0–46.6%. Yellow Dodola reduce disease incidence with 26.7–60%. The dried plant residue incorporated at different level influence fusarium wilt. The application of Holleta-1 dried plant residue at 20–100 g/kg of infested soil reduced disease incidence in the range 20.0–26.7%. The results imply the potential of using B. carinata green manure and dried plant residue as cultural management components in chickpea fusarium wilt disease management.     

Plant regeneration from stem cortex expiant and protoplast cultures of Brassica juncea (mustard)

Summary Plant regeneration from stem cortex explants of 13 genotypes of Brassica juncea was assessed. Regeneration was strongly affected by genotype, as up to 50.6 shoots were produced per 100 calli of the most responsive line (Blaze), whereas no shoots were obtained from less responsive lines (Zeml, Vniimk351). Blaze was chosen for B. juncea stem cortex protoplast isolation. After one week of culture, 11–14% of the cells had divided, and about 0.002% produced 1–2 mm colonies within 6 weeks. Up to 7% of these colonies gave rise to shoots upon transfer to plant regeneration medium.Abbreviations 2,4-D 2,4-dichlorophenoxyaceticacid - BAP 6-benzylaminopurine - FDA fluorescein diacetate - IAA indole acetic acid - IBA indole butyric acid - 2ip 2-isopentenyl-adenine - NAA 1-naphthaleneacetic acid - TD thidiazuron     

Effect of ‘Mixtalol’ on growth,yield and yield components of Indian mustard (Brassica juncea)

Mixtalol (a mixture of long chain aliphatic alcohols varying in chain length from C₂₄ to C₃₂) applied to Brassica juncea plants as foliar spray caused an increase in secondary and tertiary branching with consequent enhancement in seed yield through increased number of inflorescences and siliquae per plant. The percentage of immature siliquae and shattering of siliquae decreased with this treatment. Mixtalol increased total dry matter of plants, partitioning coefficient and harvest index. The contents of starch, protein and oil were also higher in seeds from Mixtalol treated plants.     

Improving nitrogen fixation in legumes by plant breeding; the relevance of host selection experiments in red clover (Trifolium pratense L.) and subterranean clover (T. subterraneum L.)

Summary This paper reviews (i) basic studies on the genetics of symbiosis in red clover (a self-sterile species) and subterranean clover (cleistogamous) and (ii) work on selection and plant breeding to increase nitrogen fixation in these hosts.Symbiotic effectiveness in red clover is influenced by many major and minor genes. The highly effective phenotype is inherited in a complex manner associated with early nodulation and the formation of large amounts of persistent bacteroid-containing tissue. Lines bred to fix more nitrogen with one strain ofRhizobium trifolii do so with most but not all other strains examined. They also show slightly increased vigour when grown on nitrate. The highly effective response is correlated with abundant nodulation and an early flowering habit, the evidence from breeding studies indicating that this correlation is not absolute. Normally effective and highly effective nodules have the same specific nitrogenase activities. The expression of the highly effective response is relatively little affected by environmental factors (temperature, light intensity, day length, supplementary carbon-di-oxide). Inbreeding substantially degrades the symbiotic response.Heterosis is shown in crosses between cultivars of subterranean clover but otherwise selection to increase effectiveness in this host was unsuccessful.The relevance of these results (and their physiological aspects) for the improvement of grain legumes is discussed.     

Interspecific hybridization of red clover (Trifolium pratense L.) with T. sarosiense Hazsl. Using in vitro embryo rescue

Summary Interspecific hybrid clover plants from the cross Trifolium sarosiense Hazsl. X T. pratense L. were obtained in the present investigation. Immature hybrid embryos were excised aseptically from the pistillate parent, T. sarosiense (2 n = 48), and cultured in vitro prior to in situ abortion. Agar-solidified nutrient media modified from that developed previously for tissue and cell cultures of red clover (2 n = 14) were used for embryo rescue.The heart shaped embryos obtained were cultured for 8 to 14 days on a medium containing a high level of sucrose, a moderate level of auxin, and low cytokinin activity. Viable embryos were then transferred to a standard medium with low auxin and moderate cytokinin levels for the direct germination of shoots. Some embryos produced only callus. Plants were regenerated from callus using an alternate culture scheme. Hybrid shoot numbers were increased on a low auxin, high cytokinin medium and subsequently rooted before transfer to soil in the greenhouse.About 10% of the hybrid embryos were rescued using the optimal culture sequence. Five full-sib families of the F₁ hybrid were successfully grown to maturity. Root-tip cells of hybrid plants possessed the expected somatic chromosome number of 31. The genetically determined leaf-mark trait carried by the staminate parent and the rhizomatous root habit of the pistillate parent were expressed in hybrid plants.The investigations reported in this paper (No. 81-3-151) were performed in connection with projects of the Kentucky Agricultural Experiment Station and the paper is published with the approval of the Director     

In vivo estrogenic comparisons of Trifolium pratense (red clover) Humulus lupulus (hops), and the pure compounds isoxanthohumol and 8-prenylnaringenin

The lack of a safe and reliable alternative to hormone therapy (HT) for treating menopausal symptoms underscores the need for alternative therapies. OBJECTIVE: The purpose of this study was to assess the in vivo estrogenic effects of the botanical dietary supplements Trifolium pratense (red clover) and Humulus lupulus (hops), and two compounds obtained from H. lupulus, isoxanthohumol and 8-prenylnaringenin (8-PN) using the ovariectomized uterotrophic adult rat model. A H. lupulus extract and a 30% isoflavone extract of T. pratense were tested at three escalating doses as was one dose of isoxanthohumol for 21d. 8-Prenylnaringenin, the major estrogen in H. lupulus, was also tested at three relevant escalating doses. In order to determine the in vivo metabolism of 8-PN, the major phases I and II metabolites were also identified. The primary outcome measure, uterus weight gain, indicated that H. lupulus and T. pratense did not have an estrogenic effect on the uterus, and none of the secondary outcome measures were positive. In contrast, there was a clear dose response when 8-PN was evaluated where the middle and high doses of 8-PN were active. 8-Prenylnaringenin in rat plasma, liver, and mammary gland was measured and the major phases I and II 8-PN metabolites were detected. Our findings suggest that while both the H. lupulus and T. pratense extracts do not have an effect on the rat uterus, 8-PN at equivalent doses to those previously used in humans did have an effect, and may therefore have a deleterious effect in women.     

Research on antioxidant effects and estrogenic effect of formononetin from Trifolium pratense (red clover)

Antioxidant and estrogenic effects of formononetin on ovariectomized mice have been investigated in the present study. The adult female Kunming mice were divided into 5 groups: sham-operated group, ovariectomized group, stilbestrol replacement therapy group (0.20 mg/kg day), low-dose formononetin group (0.05 g/kg day) and high-dose formononetin group (0.5 g/kg day). The mice in the latter 4 groups were ovariectomized. The drug was given by oral administration for 6 months. Estrogenic effect was determined by the change of uterine weight, and oxidant effects were determined by the content of SOD, GSH-Px, CAT and MDA. The intake of formononetin increased the uterine weight of the mice significantly as well as the content of SOD, GSH-Px, CAT, and reduced MDA in body. Formononetin had obvious antioxidant effects and estrogenic effect, and the estrogenic effect was not dosage-related.     

长期双季稻绿肥轮作对水稻产量及稻田土壤有机质的影响

以中国农业科学院红壤实验站1982年布置的长期定位试验为研究对象,分析了长期双季稻绿肥轮作体系下水稻产量变化趋势、稻田土壤有机质变化特征及土壤活性有机质组成。结果表明,绿肥作物与双季稻轮作种植后,水稻产量显著高于冬闲对照,绿肥作物紫云英、油菜和黑麦草处理年平均水稻产量（1982-2008）分别为10.8 t?hm-2?a-1,10.2 t?hm-2?a-1和10.0 t?hm-2?a-1,较冬闲对照分别提高27.2%,20.5%和18.1%。试验前期（1982-1993）种植绿肥作物各处理之间水稻产量无显著差异,试验开展11年后（1994-2008）种植紫云英处理水稻产量显著高于油菜和黑麦草处理。长期双季稻绿肥轮作土壤有机质随年份显著增加,双季稻紫云英轮作土壤有机质积累速度最快,年增加0.31 g?kg-1,双季稻黑麦草次之,土壤有机质年增加0.28g?kg-1,双季稻油菜轮作土壤有机质年增加0.26g?kg-1。种植绿肥作物紫云英稻田土壤活性有机质显著高于其它处理。种植绿肥作物各处理土壤有机碳、全氮、土壤微生物量碳和土壤微生物量氮含量均显著高于冬闲对照。其中黑麦草和紫云英处理土壤微生物量碳含量及微生物熵显著高于油菜和冬闲对照处理。在湘南红壤丘陵双季稻区,种植绿肥作物对提高水稻产量、增加土壤有机质、提高土壤有机质活性具有重要意义,绿肥选择上以紫云英对水稻产量和稻田土壤培肥综合效果最好。     

Fatty acid profiles of monofloral clover beebread and pollen and proteomics of red clover (Trifolium pratense) pollen

Fatty acids were identified in monofloral beebread (BB) and bee pollen (BP) loads collected from Trifolium pratense L. A gas chromatography method was used to identify and quantify fatty acids: Thirty-five fatty acids were identified in BB and 42 in BP. A high amount of the healthy n-3 fatty acids was found. The ratio of polyunsaturated fatty acids n-3 to n-6 reached a value of 8.42 and 3.35 in the latter products. The proteomic analysis also was performed on the manually collected T. pratense pollen, and the most abundant protein groups were subjected to mass spectrometry analysis. Proteins identified in T. pratense pollen are involved in the main cellular functions (cell membrane formation, organelles traffic, and mainly metabolic processes). Because of the composition of fatty acids in BB and BP and a variety of proteins present in pollen, these products are considered to be favorable for human nutrition and health.     

Effect of NPK on growth and nitrogen fixation of Sesbania rostrata as a green manure for lowland rice (Oryza sativa L.)

The stem-nodulating tropical legume Sesbania rostrata is a promising green manure species for low input rice-farming systems in lowland areas. However, its success as biofertilizer depends on its biomass production and N₂ fixation. Nutrient imbalances and soils low in available nutrients can considerably affect biofertilizer production. Use of mineral N, P, and K fertilizers in growing S. rostrata as biofertilizer for lowland rice was therefore evaluated in pot experiments, and in the fields in Central Luzon, Philippines. Two soils low in Olsen P (3–7.3 mg kg^–1) and exchangeable K (0.05–0.08 meq 100g^-1) were used. Increasing amounts of N (0, 10, 20, 30, and 40 mg kg^-1), P (0, 50, and 100 mg kg^-1), and K (0, 100, 200, and 300 mg kg^-1) were applied to S. rostrata grown in the greenhouse, whereas small amounts of N, P, and K fertilizers (30, 15, and 33 kg ha^-1, respectively) were applied in the field.Mineral N application depressed nodulation and N₂ fixation in roots. It however, stimulated nodulation and N₂ fixation in stems. Applying 30 kg N ha^-1 as urea increased total N accumulation by S. rostrata and yield of the subsequent rice crop (IR64). Applied P and K both stimulated growth, nodulation, and N₂ fixation of S. rostrata. Nitrogen accumulation in P- and K-fertilized S. rostrata was about 40% higher than that in nonfertilized green manure. Thus integration of mineral N, P, and K fertilizers in a green manure-based rice-farming system can considerably improve biofertilizer production and increase rice grain yield.     

Effects of organic amendments to soil on the rhizosphere microflora of antirrhinum infected withVerticillium dahliae Kleb.

Summary Organic (e.g. chitin, green manure, cellulose) amendments to soil induced quantitative and qualitative changes in the rhizosphere microflora of antirrhinum plants infected withVerticillium dahliae Kleb. Whereas reduction in disease severity occurred with chitin and green manure amendments, an increase in disease severity was observed with the application of cellulose. The reduction of the disease severity with chitin and green manure may be correlated with the increased population of actinomycetes in the antirrhinum rhizosphere.