Phosphorus nutrition and tolerance of cotton to water stress: II. Water relations, free and bound water and leaf expansion rate

In previous experiments, increased leaf-Phosphorus (P) content with increasing P supply enhanced the individual leaf expansion and water content of fresh cotton leaves in a severely drying soil. In this paper, we report on the bulk water content of leaves and its components, free and bound water, along with other measures of plant water status, in expanding cotton leaves of various ages in a drying soil with different P concentrations. The bound water in living tissue is more likely to play a major role in tolerance to abiotic stresses by maintaining the structural integrity and/or cell wall extensibility of the leaves, whilst an increased amount of free water might be able to enhance solute accumulation, leading to better osmotic adjustment and tolerance to water stress, and maintenance of the volumes of sub-cellular compartments for expansive leaf growth. There were strong correlations between leaf-P%, leaf water (total, free and bound water) and leaf expansion rate (LER) under water stress conditions in a severely drying soil. Increased soil-P enhanced the uptake of P from a drying soil, leading to increased supply of osmotically active inorganic solutes to the cells in growing leaves. This appears to have led to the accumulation of free water and more bound water, ultimately leading to increased leaf expansion rates as compared to plants in low P soil under similar water stress conditions. The greater amount of bound and free water in the high-P plants was not necessarily associated with changes in cell turgor, and appears to have maintained the cell-wall properties and extensibility under water stressed conditions in soils that are nutritionally P-deficient.

A promising and simple method to quantify soil/manure mixing on beef feedlot pens

On beef cattle feed pen surfaces, fresh and decayed manure is mixed with base rock or soil (base). Quantifying this mixing has beneficial applications for aspects including nutrient and greenhouse gas budgeting. However, no practical methods exist to quantify mixing. We investigated if measuring element concentrations in: (A) fresh manure, (B) base material, and (C) pen manure offers a promising method to quantify manure/base mixing on pen surfaces. Using three operational beef feedlots as study sites, we targeted carbon (C), and silicon (Si), which are the two most abundant and easily measurable organic and inorganic elements. Our results revealed that C concentrations were strongly (>15 times) and significantly (P < 0.05) higher whereas Si concentrations strongly (>10 times) and significantly (P < 0.01) lower in fresh manure than base material at all three sites. These relative concentrations were not significantly impacted by manure decay, as determined by an 18-week incubation experiment. This suggested that both of these elements are suitable markers for quantifying base/manure mixing on pens. However, due to the chemical change of manure during decay, C was shown to be an imprecise marker of base/manure mixing. By contrast, using Si to estimate base/manure mixing was largely unaffected by manure decay. These findings were confirmed by measuring C and Si concentrations in stockpiled pen surface manure from one of the sites. Using Si concentrations is a promising approach to quantify base/manure mixing on feed pens given that this element is abundantly concentrated in soils and rocks.

Temporal variation in arthropod sampling effectiveness: The case for using the beat sheet method in cotton

Predatory insects and spiders are key elements of integrated pest management (IPM) programmes in agricultural crops such as cotton. Management decisions in IPM programmes should to be based on a reliable and efficient method for counting both predators and pests. Knowledge of the temporal constraints that influence sampling is required because arthropod abundance estimates are likely to vary over a growing season and within a day. Few studies have adequately quantified this effect using the beat sheet, a potentially important sampling method. We compared the commonly used methods of suction and visual sampling to the beat sheet, with reference to an absolute cage clamp method for determining the abundance of various arthropod taxa over 5 weeks. There were significantly more entomophagous arthropods recorded using the beat sheet and cage clamp methods than by using suction or visual sampling, and these differences were more pronounced as the plants grew. In a second trial, relative estimates of entomophagous and phytophagous arthropod abundance were made using beat sheet samples collected over a day. Beat sheet estimates of the abundance of only eight of the 43 taxa examined were found to vary significantly over a day. Beat sheet sampling is recommended in further studies of arthropod abundance in cotton, but researchers and pest management advisors should bear in mind the time of season and time of day effects.

Vermiculite improves early development and survival of low chill stone-fruit embryos rescued in vitro

There are many reports of efficient embryo germination and the method has been optimized to suit subtropical low chill genotypes. However the subsequent growth, vigor, and ability of germinated embryos to develop and survive acclimatization is rarely reported. Many germinated embryos do not survive acclimatization, develop slowly, or fail to develop normally. Methods to improve plant development from in vitro embryo cultures are needed to improve the number of plants that survive to be useful in breeding programs. This paper describes an improved method of embryo rescue that significantly increases embryo shoot and root development that leads to increased plant survival. Four treatments: Woody Plant Media (WPM) solidified with agar, vermiculite with liquid WPM, vermiculite with WPM plus agar, and conventional stratification, were evaluated for embryo growth and subsequent plantlet development and survival for two low-chill peach and one low-chill nectarine cultivar. Highly significant improvements were found for shoot and root development of seedlings germinated in vermiculite based media compared to embryos germinated in conventional agar-based media. Vermiculite with WPM and agar improved plantlet growth subsequent to in vitro culture and significantly increased survival of germinated embryos resulting in more plants reaching the field.

Campylobacter jejuni and Campylobacter coli Genotyping by High-Resolution Melting Analysis of a flaA Fragment.

The highly variable flagellin-encoding flaA gene has long been used for genotyping Campylobacter jejuni and Campylobacter coli. High-resolution melting (HRM) analysis is emerging as an efficient and robust method for discriminating DNA sequence variants. The objective of this study was to apply HRM analysis to flaA-based genotyping. The initial aim was to identify a suitable flaA fragment. It was found that the PCR primers commonly used to amplify the flaA short variable repeat (SVR) yielded a mixed PCR product unsuitable for HRM analysis. However, a PCR primer set composed of the upstream primer used to amplify the fragment used for flaA restriction fragment length polymorphism (RFLP) analysis and the downstream primer used for flaA SVR amplification generated a very pure PCR product, and this primer set was used for the remainder of the study. Eighty-seven C. jejuni and 15 C. coli isolates were analyzed by flaA HRM and also partial flaA sequencing. There were 47 flaA sequence variants, and all were resolved by HRM analysis. The isolates used had previously also been genotyped using single-nucleotide polymorphisms (SNPs), binary markers, CRISPR HRM, and flaA RFLP.flaA HRM analysis provided resolving power multiplicative to the SNPs, binary markers, and CRISPR HRM and largely concordant with the flaA RFLP. It was concluded that HRM analysis is a promising approach to genotyping based on highly variable genes.

Enhancing somatic embryogenesis in avocado (Persea americana Mill.) using a two-step culture system and including glutamine in the culture medium

The development of a more efficient in vitro regeneration system for somatic embryos (SEs) of avocado (Persea americana) would facilitate the development of new superior cultivars for this valuable horticultural crop. In this study, we report a new and efficient method for maintenance and regeneration of avocado SEs. Avocado SEs of four cultivars remained healthy and viable in vitro for 11 months on a medium used for mango somatic embryogenesis, compared with 3-4 months on Murashige and Skoog medium. Various supplements and media modifications were investigated to improve the low conversion rate of regenerated plants from avocado SEs reported previously. The one-step system for regeneration of white-opaque somatic embryos (WOSEs) used solid medium only over a period of 12-14 weeks (sub-culturing every 6 weeks). Addition of praline and glutamine improved the total regeneration from 0 to 17.5% and 10.5%, and plant/shoot recovery from 0 to 12.5% and 5%, respectively. A two-step culture system involving the transfer of WOSEs of cultivar 'Reed' after 6 weeks on solid to liquid medium for 12-15 days as an intermediate step, followed by subculturing again onto solid medium for 6 weeks improved total regeneration to 29% and plant/shoot recovery to 18.3 from 0% when regenerated by subculturing on solid medium only. Supplementation with proline in the solid as well as liquid medium in the two-step culture system at 0.4 g/L increased total regeneration to 35% and plant/shoot recovery to 20%. We were able to achieve highest regeneration using glutamine at 1 g/L in the two-step culture system in terms of both total regeneration (58.3%, including 43.3% bipolar regeneration) and plant/shoot recovery (36.7%) rates, which were significantly higher than in any other treatment investigated. (C) 2013 Elsevier B.V. All rights reserved.

Welfare outcomes for 3- and 6-month-old beef calves in a tropical environment castrated surgically or by applying rubber rings

Castration of cattle using rubber rings is becoming increasingly popular due to the perceived ease of the procedure and greater operator safety when compared with surgical castration. Few comparative studies have investigated the effects of different castration methods and calf age on welfare outcomes, particularly in a tropical environment. Thirty Belmont Red (a tropically adapted breed), 3-month-old (liveweight 71–119 kg) and 30, 6-month-old (liveweight 141–189 kg) calves were assigned to a two age × three castration (surgical, ring and sham) treatment factorial study (Surg3, Surg6, Ring3, Ring6, Sham3 and Sham6, n = 10 for each treatment group). Welfare outcomes were assessed post-castration using: behaviour for 2 weeks; blood parameters (cortisol and haptoglobin concentrations) to 4 weeks; wound healing to 5 weeks; and liveweights to 6 weeks. More Surg calves struggled during castration compared with Sham and Ring (P < 0.05, 90 ± 7% vs. 20 ± 9% and 24 ± 10%) and performed more struggles (1.9 ± 0.2, 1.1 ± 0.3 and 1.1 ± 0.3 for Surg, Sham and Ring, respectively), suggesting that surgical castration caused most pain during performance of the procedure. A significant (P < 0.05) time × castration method × age interaction for plasma cortisol revealed that concentrations decreased most rapidly in Sham; the Ring6 calves failed to show reduced cortisol concentrations at 2 h post-castration, unlike other treatment groups. By 7 h post-castration, all treatment groups had similar concentrations. A significant (P < 0.01) interaction between time and castration method showed that haptoglobin concentrations increased slightly to 0.89 and 0.84 mg/mL for Surg and Ring, respectively over the first 3 days post-castration. Concentrations for Surg then decreased to levels similar to Sham by day 21 and, although concentrations for Ring decreased on day 7 to 0.76 mg/mL, they increased significantly on day 14 to 0.97 mg/mL before reducing to concentrations similar to the other groups (0.66 mg/mL) by day 21. Significantly (P < 0.05) more of the wounds of the 3-month compared with the 6-month calves scored as ‘healed’ at day 7 (74% vs. 39%), while more (P = 0.062) of the Surg than Ring scored as ‘healed’ at day 21 (60% vs. 29%). At day 14 there were significantly (P < 0.05) fewer healed wounds in Ring6 compared with other treatment groups (13% vs. 40–60%). Liveweight gain was significantly (P < 0.05) greater in 3-month (0.53 kg/day) than in 6-month calves (0.44 kg/day) and in Sham calves (P < 0.001, 0.54 kg/day), than in Ring (0.44 kg/day) and Surg (0.48 kg/day) calves. Overall, welfare outcomes were slightly better for Surg than Ring calves due to reduced inflammation and faster wound healing, with little difference between age groups.

Welfare outcomes for 3- and 6-month-old beef calves in a tropical environment castrated surgically or by applying rubber rings

Castration of cattle using rubber rings is becoming increasingly popular due to the perceived ease of the procedure and greater operator safety when compared with surgical castration. Few comparative studies have investigated the effects of different castration methods and calf age on welfare outcomes, particularly in a tropical environment. Thirty Belmont Red (a tropically adapted breed), 3-month-old (liveweight 71–119 kg) and 30, 6-month-old (liveweight 141–189 kg) calves were assigned to a two age × three castration (surgical, ring and sham) treatment factorial study (Surg3, Surg6, Ring3, Ring6, Sham3 and Sham6, n = 10 for each treatment group). Welfare outcomes were assessed post-castration using: behaviour for 2 weeks; blood parameters (cortisol and haptoglobin concentrations) to 4 weeks; wound healing to 5 weeks; and liveweights to 6 weeks. More Surg calves struggled during castration compared with Sham and Ring (P < 0.05, 90 ± 7% vs. 20 ± 9% and 24 ± 10%) and performed more struggles (1.9 ± 0.2, 1.1 ± 0.3 and 1.1 ± 0.3 for Surg, Sham and Ring, respectively), suggesting that surgical castration caused most pain during performance of the procedure. A significant (P < 0.05) time × castration method × age interaction for plasma cortisol revealed that concentrations decreased most rapidly in Sham; the Ring6 calves failed to show reduced cortisol concentrations at 2 h post-castration, unlike other treatment groups. By 7 h post-castration, all treatment groups had similar concentrations. A significant (P < 0.01) interaction between time and castration method showed that haptoglobin concentrations increased slightly to 0.89 and 0.84 mg/mL for Surg and Ring, respectively over the first 3 days post-castration. Concentrations for Surg then decreased to levels similar to Sham by day 21 and, although concentrations for Ring decreased on day 7 to 0.76 mg/mL, they increased significantly on day 14 to 0.97 mg/mL before reducing to concentrations similar to the other groups (0.66 mg/mL) by day 21. Significantly (P < 0.05) more of the wounds of the 3-month compared with the 6-month calves scored as ‘healed’ at day 7 (74% vs. 39%), while more (P = 0.062) of the Surg than Ring scored as ‘healed’ at day 21 (60% vs. 29%). At day 14 there were significantly (P < 0.05) fewer healed wounds in Ring6 compared with other treatment groups (13% vs. 40–60%). Liveweight gain was significantly (P < 0.05) greater in 3-month (0.53 kg/day) than in 6-month calves (0.44 kg/day) and in Sham calves (P < 0.001, 0.54 kg/day), than in Ring (0.44 kg/day) and Surg (0.48 kg/day) calves. Overall, welfare outcomes were slightly better for Surg than Ring calves due to reduced inflammation and faster wound healing, with little difference between age groups.