Development and use of microsatellite markers for genetic diversity analysis of cañahua (<Emphasis Type="Italic">Chenopodium pallidicaule</Emphasis> Aellen)

Cañahua (Chenopodium pallidicaule Aellen) is a poorly studied, annual subsistence crop of the high Andes of South America. Its nutritional value (high in protein and mineral content) and ability to thrive in harsh climates make it an important regional food crop throughout the Andean region. The objectives of this study were to develop genetic markers and to quantify genetic diversity within cañahua. A set of 43 wild and cultivated cañahua genotypes and two related species (Chenopodium quinoa Willd. and Chenopodium petiolare Kunth) were evaluated for polymorphism using 192 microsatellite markers derived from random genomic cañahua sequences produced by 454 pyrosequencing of cañahua genomic DNA. Another 424 microsatellite markers from C. quinoa were also evaluated for cross-species amplification and polymorphism in cañahua. A total of 34 polymorphic microsatellite marker loci were identified which detected a total of 154 alleles with an average of 4.5 alleles per marker locus and an average heterozygosity value of 0.49. A cluster analysis, based on Nei genetic distance, clearly separated from wild cañahua genotypes from the cultivated genotypes. Within the cultivated genotypes, subclades were partitioned by AMOVA analysis into six model-based clusters, including a subclade consisting sole of erect morphotypes. The isolation by distance test displayed no significant correlation between geographic collection origin and genotypic data, suggesting that cañahua populations have moved extensively, presumably via ancient food exchange strategies among native peoples of the Andean region. The molecular markers reported here are a significant resource for ongoing efforts to characterize the extensive Bolivian and Peruvian cañahua germplasm banks, including the development of core germplasm collections needed to support emerging breeding programs.     

Relationship between silver scurf levels on seed and progeny tubers from successive generations of potato seed

The level of silver scurf on potato seed tubers on successive generations of potato seed tubers and their progeny tubers was investigated during 3 years in the field. The objective was to determine the importance of seed-borne inoculum on silver scurf development on the subsequent progeny tubers. Silver scurf incidence and severity increased with each generation. Coefficients of determination for disease levels among generations were significant and ranged from 0.89 to 0.97, indicating that seed tuber source accounted for a large proportion of silver scurf on progeny seed tubers. Incidence and severity of silver scurf also increased with decreasing time periods between potato crops in the field. In a field near Paterson, WA, where potatoes had not been previously grown, the severity of silver scurf increased on progeny tubers of cvs Russet Norkotah, Ranger Russet, and Shepody as disease severity increased on seed tubers of successive generations. Disease severity index significantly increased as disease incidence increased. The relationship between the two was best described using a curvilinear regression model.     

Deficient,Adequate and Excess Nitrogen Levels Established in Hydroponics for Biotic and Abiotic Stress-Interaction Studies in Potato

Application of mineral nutrients to improve plant health is critical to providing food and fiber. Potato (Solanum tuberosum L.) is among the most responsive species to N application but is susceptible to biotic and abiotic stresses. Nitrogen-stress relationships are generally evaluated indirectly during field fertility trials designed for collection of agronomic data. A controlled environment system would benefit understanding N nutrition-stress interactions. Environmental chamber hydroponics allows strict experimental control of temperature, humidity, light, etc. while maintaining specific N nutrition. Potatoes were grown at various N levels. Plant biomass had a curvilinear relationship with N supply, peaking at 80 mg N L^?1. Root or shoot N regressed against solution N produced significant, predictable relationships (R² > 0.98 for each). Such relationships provide a basis to conduct controlled N nutrition-abiotic-biotic stress studies that could delineate among stress factors. This work establishes the specific ranges of N needed to complete such research.     

Ability of intravaginal progesterone inserts and melengestrol acetate to induce estrous cycles in postpartum beef cows

Postpartum anestrous interval in beef cows is a major factor contributing to reproductive failure during a defined breeding season. Our objectives were to determine the ability of a controlled internal drug-releasing device (CIDR, 1.9 g of progesterone), a normal dose of melengestrol acetate (MGA, 0.5 mg x cow(-1) x d(-1)), or a high dose of MGA (4.0 mg x cow(-1) x d(-1)) to induce ovulation and to eliminate short estrous cycles. Multiparous beef cows (n = 100) were equally assigned to one of four treatments: CIDR, normal MGA, high MGA, or control by age, days postpartum, body condition, and body weight. All cows were fed carrier (0.9072 kg x cow(-1) x d(-1)) with (normal MGA, 0.55 mg/kg; high MGA, 4.41 mg/kg) or without MGA for 7 d (d -6 to 0). On d -6, CIDR were inserted and then removed on d 0. Estrous behavior was monitored continuously from d -6 until 29 using HeatWatch electronic mount detectors. Blood was collected on d -13, and three times weekly from d -6 to 29. Treatment influenced (P = 0.03) the percentage of cows that were detected in standing estrus. Beginning on d 2, more CIDR-treated cows had exhibited standing estrus compared with high MGA-treated or control cows, but CIDR- and normal MGA-treated cows did not differ. The percentage of CIDR-treated cows that had ovulated was greater (P < 0.05) than the percentage of normal MGA-treated, high MGA-treated, or control cows beginning on d 4. The percentage of cows that exhibited standing estrus before the first postpartum ovulation (CIDR = 65%, normal MGA = 57%, high MGA = 35%, control = 30%) did not differ (P = 0.09) among treatments. Luteal life span following the first ovulation postpartum and the percentage of cows with a normal luteal life span (i.e., progesterone > 1 ng/mL for > or = 10 d) was greater (P < 0.01) in CIDR-treated cows (14.0 +/- 0.8 d; 20/20, 100%) compared with normal MGA-treated (6.2 +/- 1.0 d; 3/13, 23%), high MGA-treated (9.6 +/- 1.0 d; 8/14, 57%), or control cows (6.1 +/- 0.9 d; 4/17, 24%), and greater (P < 0.03) in high MGA-treated cows than in normal MGA-treated or control cows. In the present study, treatment of early postpartum suckled beef cows with CIDR induced ovulation and initiated estrous cycles with a normal luteal life span in more cows than did treatment with MGA. Treatment with MGA (normal or high dose) did not induce ovulation earlier than in control cows, but a high dose of MGA increased the percentage of cows with normal luteal life spans following the first ovulation postpartum.     

Anthelmintic paraherquamides are cholinergic antagonists in gastrointestinal nematodes and mammals

Oxindole alkaloids in the paraherquamide/marcfortine family exhibit broad-spectrum anthelmintic activity that includes drug-resistant strains of nematodes. Paraherquamide (PHQ), 2-deoxoparaherquamide (2DPHQ), and close structural analogs of these compounds rapidly induce flaccid paralysis in parasitic nematodes in vitro, without affecting adenosine triphosphate (ATP) levels. The mechanism of action of this anthelmintic class was investigated using muscle tension and microelectrode recording techniques in isolated body wall segments of Ascaris suum. None of the compounds altered A. suum muscle tension or membrane potential. However, PHQ blocked (when applied before) or reversed (when applied after) depolarizing contractions induced by acetylcholine (ACh) and the nicotinic agonists levamisole and morantel. These effects were mimicked by the nicotinic ganglionic blocker mecamylamine, suggesting that the anthelmintic activity of PHQ and marcfortines is due to blockade of cholinergic neuromuscular transmission. The effects of these compounds were also examined on subtypes of human nicotinic ACh receptors expressed in mammalian cells with a Ca2+ flux assay. 2DPHQ blocked nicotinic stimulation of cells expressing alpha3 ganglionic (IC50 approximately 9 microm) and muscle-type (IC50 approximately 3 microm) nicotinic cholinergic receptors, but was inactive at 100 microm vs. the alpha7 CNS subtype. PHQ anthelmintics are nicotinic cholinergic antagonists in both nematodes and mammals, and this mechanism appears to underlie both their efficacy and toxicity.     

Kepler-16: a transiting circumbinary planet

We report the detection of a planet whose orbit surrounds a pair of low-mass stars. Data from the Kepler spacecraft reveal transits of the planet across both stars, in addition to the mutual eclipses of the stars, giving precise constraints on the absolute dimensions of all three bodies. The planet is comparable to Saturn in mass and size and is on a nearly circular 229-day orbit around its two parent stars. The eclipsing stars are 20 and 69% as massive as the Sun and have an eccentric 41-day orbit. The motions of all three bodies are confined to within 0.5° of a single plane, suggesting that the planet formed within a circumbinary disk.     

Development of real-time diagnostic assays specific for Mycoplasma mycoides subspecies mycoides Small Colony

Rapid and specific detection of Mycoplasma mycoides subsp. mycoides Small Colony (M. mycoides SC) is important for the effective control of contagious bovine pleuropneumonia. Although the United States has been free of this disease for over 100 years, it is necessary to develop modern diagnostic assays that are sensitive and specific for biological agents that would affect the US agricultural industry following accidental or intentional introduction into the US agricultural population. With this aim in mind, we have identified M. mycoides SC-specific genetic loci and developed TaqMan-based PCR assays for the detection of M. mycoides SC. The TaqMan assay allows for real-time detection of specific, amplified PCR products using portable equipment, enabling testing to be performed in the field. These assays are specific for M. mycoides SC, failing to amplify DNA from other organisms belonging to the M. mycoides cluster or two phylogenetically unrelated bovine mycoplasma species. Standard curves were drawn based on the linear relationships measured between the threshold fluorescence (C(T)) values and a measured quantity of genomic DNA. M. mycoides SC was successfully detected in bronchoalveolar lavage samples obtained from experimentally infected cattle. These TaqMan-based real-time PCR assays will allow for the rapid and specific detection of M. mycoides SC.