Effects of tractor wheeling on root morphology and yield of lucerne (Medicago sativa L.)

The purpose of this study was to determine the effect of soil compaction on the herbage yield and root growth of lucerne ( Medicago sativa L.). A field experiment was conducted on a silty loam Mollic Fluvisols soil in 2003–2006. Herbage yield and root morphology, in terms of root length density, mean root diameter, specific root length and distribution of dry matter (DM) in roots, were measured. Four compaction treatments were applied three times annually by tractor using the following number of passes: control without experimental traffic, two passes, four passes and six passes. The tractor traffic changed the physical properties of the soil by increasing bulk density and penetration resistance. Soil compaction also improved its water retention properties. These changes were associated with changes in root morphology and distribution of the DM in roots. Soil compaction resulted in higher proportions of the DM in roots, especially in the upper, 0–10 cm, soil horizon. Decreases in the root length density were observed in a root diameter range of 0·1–1·0 mm. It was also found that roots in a more compacted soil were significantly thicker. An effect of the root system of lucerne on soil compaction was observed. The root system of lucerne decreased the effects of soil compaction that had been recorded in the first and the second year of the experiment. An increase in the number of passes resulted in a decrease in the DM yield of herbage in the second and third harvests each year.     

花生体细胞胚胎诱导和植株再生研究

以花生成熟种胚为外植体,在ＭＳ附加不同激素（２０ｍｇ／Ｌ２,４－Ｄ或５ｍｇ／Ｌ Ｐｉｃｌｏｒａｍ）的培养基上黑暗培养,胚性愈伤组织发生率和产胚量无明显差异,在ＭＳ＋５ｍｇ／Ｌ Ｐｉｃｌｏｒａｍ培养基上,胚性愈伤组织发生率和产胚量和品种类型有关,珍珠豆型花生品种胚性愈伤组织发生率和产胚量高于普通型花生品种。体胚在ＭＳ＋１０ｍｇ／Ｌ ＢＡ培养基上苗再生达３６．３％－７７．８％,再生小苗在ＭＳ＋０．３ｍ     

Effects of sulphur supply on the morphology of shoots and roots of alfalfa (Medicago sativa L.)

A sulphur (S)‐deficient top soil was used in a pot experiment to investigate the effect of S supply on shoot and root growth and development in alfalfa (Medicago sativa L.). The treatments consisted of three rates of addition of S: 0, 20 and 40 mg kg^?1 soil and each was replicated four times. Alfalfa was harvested at 15, 30, 45, 60 and 75 d after seedling emergence. By the end of the experiment, plants with S supply had a significantly larger leaf area, heavier leaf, shoot and root dry weight per pot than controls. The effects of adding S also significantly increased plant height, basal stem diameter, chlorophyll concentration of young leaves, root length and root surface area compared with controls. The effects of S were greater on shoots than on roots. The ratio of root to shoot dry weight was 0·47 when S was supplied and 0·88 without added S, indicating that c. 0·32 and 0·47 of the total net photosynthate, produced with or without S supply, respectively, were used for the development of roots. Overall, overcoming S deficiency resulted in a significant increase in shoot and root growth.     

Callus induction and in vitro plant regeneration of rice (Oryza sativa L.) under various conditions

The objective of the present study was to develop an effective protocol for optimum callus induction and complete plant regeneration for four varieties of rice (Oryza sativa L.) i.e., Super Basmati, Basmati-370, Basmati-371 and Fakhre Malakand. Calli were induced from mature seed scutelum. The Murashige and Skoog (MS) and Chu's N6 media containing hormone 2, 4-D (2, 4-Dichlorophenoxy acetic acid) in different concentrations were used for callus induction. Fakhre Malakand produced maximum calli on N6 media containing 3 mg L(-1) 2,4-D. while other three varieties showed maximum callus induction on N6 media containing 2.5 mg L(-1) 2,4-D. N6 media was found better than MS media for callus induction. For complete plant regeneration the calli of two varieties i.e., Basmati-370 and Basmati-371 were plated on N6 media containing different concentrations of NAA (1-Naphthalene acetic acid) and BAP (6-benzyl aminopurine). The maximum regeneration frequency (%) was observed on N6 media containing NAA 1 mg L(-1) and BAP 2.5 mg L(-1). It took 27-30 days for the callus to regenerate into a complete plant. Basmati-370 produced 4-7 plantlets per callus whereas Basmati-371 produced 4-8 plantlets per callus with regeneration frequencies of 61 and 69%, respectively.     

花生细胞工程与遗传转化研究进展

花生细胞工程与遗传转化的研究对于花生品种改良、新品种繁育以及种质资源保存等具有重要意兰。对二十世纪90年代以来花生高效再生技术和遗传转化研究的现状以及发展趋势进行概述和讨论。     

In vitro reproduction in the annual pasture legumes subterranean clover (Trifolium subterraneum L.) and French serradella (Ornithopus sativus Brot.)

Pasture legumes are important components of both mixed farming rotations and permanent pastures in temperate climates. Breeding of two widely sown pasture legumes, subterranean clover (Trifolium subterraneum L.) and French serradella (Ornithopus sativus Brot.), is constrained by the long generation cycle, typically enabling only one generation per year. We hypothesized manipulation of culture medium and conditions would enable the development of a laboratory‐based protocol for in vitro reproduction in subterranean clover and French serradella. In vitro flowering and viable seed set was induced from both species. For subterranean clover, the most effective treatment was culturing on modified MS medium with 1 μm kinetin and 0·1 m sucrose under a 100 μmol m^?2 s^?1 light intensity and continuous photoperiod. For French serradella, culture on a hormone‐free B5 medium with 5 mm NH₄Cl and 0·1 m sucrose under a 100 μmol m^?2 s^?1 light intensity and 20 h photoperiod was optimum. It is expected this technique will have application in accelerating generation turnover within breeding programs, for the study of factors influencing flowering in pasture legumes, and for the propagation of valuable yet enfeebled plants such as embryo‐rescued hybrids.     

In vitro rumen methane output of grasses and grass silages differing in fermentation characteristics using the gas‐production technique (GPT)

The chemical composition of silage consumed by cattle can influence the subsequent rumen microbial fermentation patterns and methane (CH₄) emissions. The objectives of this study were to (i) evaluate the effect of ensilage on the in vitro rumen methane output of perennial ryegrass and (ii) relate the silage fermentation characteristics of grass silages with in vitro rumen methanogenesis. Three pre‐harvest herbage‐conditioning treatments and seven silage‐additive treatments were used in a laboratory‐scale silo experiment to produce a diversity of silage fermentation characteristics. Ensilage reduced (P < 0·01) the in vitro rumen CH₄ output (mL CH₄ g^?1 dry matter (DM) disappeared). This reflected differences in the direction of rumen fermentation (lower acetic (P < 0·05) and higher propionic (P < 0·001) acid proportions in volatile fatty acids) rather than major changes in the extent of in vitro rumen fermentation (i.e. mmol VFA g^?1 DM). The magnitude of the decrease in CH₄ output (mL g^?1 DM incubated) owing to ensilage increased as the extent of silage fermentation, and in particular the lactic acid concentration, increased. In contrast, among silages with relatively similar extents of silage fermentation (i.e. total fermentation products), an increase in the proportion of lactic acid in silage fermentation products led to a more extensive in vitro rumen fermentation and thus to a greater CH₄ output (mL g^?1 DM).     

Potential of Cistus ladanifer L. (rockrose) in small ruminant diets – effect of season and plant age on chemical composition,in vitro digestibility and antioxidant activity

Shrubs can provide an important contribution to the fodder resources for small ruminants in Mediterranean areas, but there is limited information on their feed value, including secondary metabolites and their seasonal differences. This study evaluated the effect of seasonal variation in chemical composition, in vitro digestibility and antioxidant activity of the aerial parts of plants of Cistus ladanifer of two age groups [young plants vs. older ones (2–6 years old)]. Aerial parts of C. ladanifer plants of both age groups were characterized by moderate cell‐wall content [321–410 g NDF kg^?1 dry matter (DM)], high levels of phenolic compounds (55·1–106 g gallic acid equivalents per kg DM) and condensed tannins (CT) (32·1–161 g kg^?1 DM), low protein content (55–100 g kg^?1 DM) and low digestibility (249–315 g of digestible organic matter per kg DM). During autumn and winter, C. ladanifer showed higher protein levels and lower cell‐wall content than in the other seasons. The highest values of phenolic compounds, CT and antioxidant activity were achieved during summer. Young plants showed higher levels of phenolic compounds during spring, summer and winter, and higher CT contents in summer (more than 54 g kg^?1 DM) compared to old plants. Aerial parts of C. ladanifer of both age groups may be used as a component of ruminant nutrition, but only as a supplement and associated with other feeding resources to complement its nutritional imbalances.     

In vitro propagation and regeneration of an industrial plant kenaf (Hibiscus cannabinus L.)

The present study report a protocol for the efficient in vitro propagation of kenaf (Hibiscus cannabinus L., an industrial crop having high cellulosic fiber content) on hormone free MS medium using the shoot apex and nodal explants. Shoot tips and nodes were isolated from 15 days old seedlings cultivated on MS medium. Different combinations and concentrations of auxin/cytokinin were used and added to the MS medium to assess the shoot and root induction of theses explants. Several subcultures were drived in order to enhance the multiplication rate. Healthy and well developed in vitro propagated shoots were transferred for acclimatization under greenhouse conditions in pots filled with different substrates (sand + compost or perlite). Our results showed that shoots could elongate and root within 4-6 weeks on MS basal medium without any callus formation. However, addition of growth regulators to the MS medium leaded to a decrease in shoot and root induction rates. Indeed, the highest shoot regeneration frequency (90.5%) was obtained on MS control medium. Elongated shoots were transferred onto the same hormone free MS medium using five subcultures where the multiplication rate reached the highest value (3.66) at the fifth and last step. The in vitro rooted plantlets were acclimatized in greenhouse and successfully transplanted to natural conditions with 70% survival.     

Seed dormancy and dormancy‐breaking methods in Leymus chinensis (Trin.) Tzvel. (Poaceae)

Leymus chinensis (Trin.) Tzvel. is a perennial grass with high productivity and forage value; however, poor stand establishment, often due to seed dormancy, limits its widespread use for forage production. To investigate the mechanism of seed dormancy and to develop effective methods of improving germination, the contribution of each part of the caryopsis to dormancy was investigated, and a number of single or combined dormancy‐breaking pre‐treatments were conducted using three seed lots. The palea, lemma, pericarp/testa, and endosperm all contributed to seed dormancy. The contribution of each part to dormancy was 23·4%, lemma; 6·2%, palea; 28·4%, pericarp/testa; and 42·0%, endosperm. Hull (palea and lemma) removal, pericarp/testa piercing, and soaking in distilled water or 30% sodium hydroxide (NaOH) significantly decreased the percentage of dormant seeds (i.e. increased germination). Treating hull‐removed and pericarp/testa‐pierced seeds with gibberellic acid (GA₃) also significantly decreased the percentage of dormant seeds. Compared with each of the single pre‐treatments, the combined pre‐treatment of pre‐soaking in water for 1 d, then 30% NaOH for 60 min and treating with 300 μm GA₃ resulted in the highest germination (89%); and seed viability was 91%.     

Influence of boron on seed yield and seed quality of organic white clover (Trifolium repens L.)

The effects of boron (B) application on flower development, nectar production, germination and seed yield were investigated in organic seed crops of white clover grown in soils with <1 mg B kg^?1. Application techniques of a liquid B product were studied in two field experiments in 2009–2011. Three rates of B application to the soil at sowing and three rates of B as a foliar application in the harvest year were compared with untreated control plots. None of the B treatments had a significant effect on seed yield, relative to the control. In one experiment, with a dense crop stand, the results indicated a small (10%) yield increase with B applied to the soil. Seed germination and B concentration of shoots, roots and seeds were unaffected by B application, although the soil concentration increased. In seeds with an average B concentration of 22 mg kg^?1, 73% germinated as normal seedlings and 14% were assessed as hard seeds; corresponding values for seeds with an average B concentration of 17 mg kg^?1 were 60 and 38% respectively. The results indicate the need for further studies of factors determining B uptake in white clover .     

Response of plant species composition,biomass production and biomass chemical properties to high N,P and K application rates in Dactylis glomerata‐ and Festuca arundinacea‐dominated grassland

Little is known about the immediate effect of high nitrogen (N), phosphorus (P) and potassium (K) application rates on sown grasslands cut twice per year. We asked how quickly plant species composition, biomass yield, biomass chemical properties and nutrient balance respond to N, P and K application. An experiment using unfertilized control, P, N, NP and NPK treatments was established on seven‐year‐old cut grassland in the Czech Republic in 2007 and monitored over four years. Annual application rates were 300 kg N ha^?1, 80 kg P ha^?1 and 200 kg K ha^?1. The immediate response of plant species composition to N application was recorded and was found to be different to the response over the four years of the study period. Highly productive grasses (Dactylis glomerata, Festuca arundinacea and Phleum pratense) were promoted by N application in 2008 and then retreated together with legumes (Medicago sativa, Trifolium pratense and Trifolium repens) in all N treatments where the expansion of perennial forbs (Urtica dioica and Rumex obtusifolius) and annual weeds (Galinsoga quadriradiata, Impatiens parviflora, Lamium purpureum and Stellaria media) was recorded. At the end of the experiment, Festuca rubra was the dominant grass in the control and P treatment, and species richness was lowest in all treatments with N application. Mean annual dry‐matter yield over all years was 3.5, 3.9, 5.8, 5.6 and 6.8 t ha^?1 in the control, P, N, NP and NPK treatments, respectively. Concentrations of N in the biomass ranged from 20.0 to 28.7 g kg^?1 in the P and N treatments; concentrations of P ranged from 3.2 to 3.7 g kg^?1 in the N and P treatments; and concentrations of K ranged from 24.1 to 34.0 g kg^?1 in the NP and NPK treatments. The N:P, N:K and K:P ratios did not correctly indicate the nutrient limitation of biomass production, which was primarily N‐limited, and K‐limitation was only recorded for high production levels in treatments with N applications. On the basis of the nutrient‐balance approach, the balanced annual application rates were estimated as 140 kg N ha^?1, 30 kg P ha^?1 and 100 kg K ha^?1. We concluded that high N, P and K application rates can very quickly and dramatically change species composition, biomass production and its chemical properties in sown cut grasslands. High N application rates can be detrimental for tall forage grasses and can support the spread of weedy species.     

Grazing behaviour,intake and performance of dairy ewes with restricted access time to berseem clover (Trifolium alexandrinum L.) pasture

The effects of restricted access time to pasture (2, 4 or 6 h d^?1; 2H, 4H or 6H) on ingestive behaviour and performance were assessed on four occasions per target grazing day (D1, initial day; D4, intermediate day; and D7, final day) in dairy ewes rotationally grazing berseem clover with a 7‐day grazing period and a 21‐day recovery period. A randomized block design with two replicates per treatment was used. All ewes were supplemented daily with 700 g per head of concentrates and 700 g per head of ryegrass‐based hay. Pasture subplot and animal group data were analysed by a factorial model including access time (AT), grazing day (D) and their interaction as fixed factors. Sward height decreased from D1 (P < 0·001) and green leaf mass from D4 (P < 0·001) onwards during the grazing period. Grazing time as a proportion of AT was higher in 2H than in 4H and 6H ewes on D1 and D4 but not on D7 (P < 0·05 for AT × D). Herbage intake rate was higher in 2H than in 4H and 6H ewes (P < 0·001). Herbage and total intakes were higher in 4H and 6H than in 2H ewes (P < 0·001), with herbage intake varying non‐linearly during the grazing period (P < 0·05). Milk yield was higher in 4H and 6H than in 2H ewes (P < 0·01). To conclude, despite the evidence of compensatory behaviour, restricting access time to 2 h d^?1 constrained intake and performance of dairy ewes rotationally grazing berseem clover.     

A simultaneous effect of selfing and Epichloë endophyte on forage,seed yield and turf characteristics of perennial ryegrass (Lolium perenne L.)

Ryegrass (Lolium perenne L.) is a putative turf and forage plant characterized as mainly outcrossing in which the extent of self‐fertilization and its consequences has not been investigated. In this study, selfed and half‐sib family seeds were obtained from ten clonally propagated ryegrass plants among which two were endophyte‐free (EF) and eight were endophyte‐infected (EI). Then, 20 seed populations plus two commercial varieties of Speedy and Vigor were sown and evaluated in a biennial experiment for their turf and forage characteristics. Selfing caused significant inbreeding depression for plant fresh and dry weight and leaf length and width in the first year for both EI and EF genotypes. In the second year, only leaf length remained depressed among these characters. Also, for the most growth, turf and reproductive traits, EI populations showed less depression. The presence of endophyte in self‐pollinated populations made an increasing effect for the number of seed per spike and days to heading, although seed weight was reduced. Some of the populations derived from self‐pollination and infected with fungal endophyte including S9 and S10 were even superior to the two commercial varieties in terms of both seed production and turf characteristics. However, for seed‐related characteristics including number of spikes per plant, seed weight and seed yield, EF populations including Speedy and Vigor were more reproductive than EI populations. It is concluded that selfing may increase the discrimination between EI and EF ryegrass plants because endophyte infection may mitigate the negative consequences of inbreeding depression in prone populations.     

Effect of depth and width of cultivation and sowing date on establishment of red clover (Trifolium pratense L.) by rotary slot‐seeding into grassland

The effect of different combinations of time of sowing and depth and width of soil cultivation using a rotary slot‐seeding machine to introduce red clover (Trifolium pratense) was determined for some characteristics of newly established red clover plants in a lowland Dactylis glomerata‐dominated grassland in the Czech Republic. Sowing was carried out on four sowing dates in one growing season: 11 May; 21 June (after the first cut); 9 August (after the second cut); and 19 September. The experimental treatments consisted of all combinations of four depths (5, 10, 15 and 20 cm) and four widths (5, 10, 15 and 20 cm) of soil cultivation. Measurements were made of number of plants, weight and height of plants, weight and height of roots. Measurements at 2 months after sowing showed fewer seedlings for the 11 May sowing than for later sowing dates. The number of red clover plants increased as the row width was increased up to 15 cm, regardless of the depth of slot. Positive effects of slot width on weight and height of plants and weight and length of roots were recorded. A highly positive correlation was found between weight of plants and weight of roots. Width of row was the key factor for the successful establishment of seedlings into the existing sward by slot‐seeding. A row width of 10 cm and depth of slot of 5 cm was considered a satisfactory combination.     

Effect of intercropping vetch (Vicia sativa L.), field pea (Pisum sativum L.) and triticale (X Triticosecale) on dry‐matter yield,nutritive and ensiling characteristics when harvested at two growth stages

This study aimed to investigate the effects of growth stage and legume proportion on the nutritive and ensiling characteristics of triticale–pea–vetch intercrop mixtures. The three‐species mixtures were sown at three seeding ratios to produce different legume proportions (low, medium and high proportions) and harvested at two growth stages. The six resulting forages were subsequently ensiled in mini‐silos. Forage dry‐matter (DM) yield, nutritive characteristics of fresh and ensiled mixtures and fermentation characteristics of silages were determined. Forage DM yield almost tripled between early‐stage harvest and late‐stage harvest. Nutritive characteristics of fresh and ensiled mixtures were affected by both growth stage and legume proportion: crude protein concentration and pepsin–cellulase digestibility decreased whereas NDF and ADF concentrations increased as growth stage advanced. Crude protein concentration and pepsin–cellulase digestibility were higher for the high‐legume treatment at both growth stages. All mixtures ensiled well, as indicated by low pH and high lactic and acetic acids contents. However, all silages had high soluble nitrogen and ammonia concentrations. This study showed that triticale–pea–vetch silages hold good potential net energy supply value for ruminants, especially at early‐stage growth and when legume proportion increases. However, protein value has to be improved by reducing protein breakdown during ensiling.     

A comparison of methods to assess the likely on‐farm value for meat production systems of pasture traits and genetic gain through plant breeding using phalaris (Phalaris aquatica L.) as an example

The emphasis plant breeders place on improving seasonal dry‐matter (DM) yield of pasture plants may increase farm profitability through greater supply of DM for livestock in critical seasons. Economic values (EV) for traits can be used to guide plant breeders when selecting ‘top’ pasture plants. Two methods of calculating economic value (EV) for seasonal DM traits were evaluated. These were based on the cost of otherwise acquiring or replacing the unit change in DM (‘replacement cost method’) and the opportunity cost of not having the unit change in DM, based on changes in livestock production either as a change in stocking rate or a change in liveweight gains of growing livestock (‘change in livestock production method’). Using barley replacement cost, the EV of a 1‐kg increase in phalaris DM on Australian sheep and beef farms ranged between AUD0·234 in summer and AUD0·303 in winter. In contrast, the EV for seasonal DM using the change in weaner beef calf liveweight gains ranged between AUD0·256 in summer and AUD0·515 in winter. The change in livestock production method highlighted in this study offers an alternative to the replacement cost method, or more detailed farm system modelling or experimentations designed to estimate pasture EV.