Sheeja George

Genetic diversity of the endangered and narrow endemic Piperia yadonii (Orchidaceae) assessed with ISSR polymorphisms

Highly endangered plants that are also narrow endemics are generally found to be genetically depauperate and thus are exceedingly susceptible to ecological and anthropological threats that can lead to their extinction. Piperia yadonii is restricted to a single California county within a biodiversity hotspot. We used nine primers to generate intersimple sequence repeat (ISSR) data to assess its genetic diversity and structure. Within each population, 99% of the loci were polymorphic, expected heterozygosity was low, and a majority of the loci were shared with few other populations. Forty percent of the total variation could be attributed to population differentiation while the rest (60%) resides within populations, and the genetic distances between populations were independent of the corresponding geographical distances. High divergence among populations is likely due to fragmentation and limited gene flow. Each population contains several private loci, and ideally, each should be protected to preserve the overall diversity of the species. Because P. yadonii currently retains a modest amount of genetic variation among individuals within populations, preserving and expanding the habitat at each site to allow natural expansion of populations would be additional strategies for its conservation before populations become too small to persist naturally.

Evolution of Bacillus subtilis to enhanced growth at low pressure: up-regulated transcription of des-desKR, encoding the fatty acid desaturase system.

The atmospheric pressure on Mars ranges from 1-10 mbar, about 1% of Earth pressure (∼1013 mbar). Low pressure is a growth-inhibitory factor for terrestrial microorganisms on Mars, and a putative low-pressure barrier for growth of Earth bacteria of ∼25 mbar has been postulated. In a previous communication, we described the isolation of a strain of Bacillus subtilis that had evolved enhanced growth ability at the near-inhibitory low pressure of 50 mbar. To explore mechanisms that enabled growth of the low-pressure-adapted strain, numerous genes differentially transcribed between the ancestor strain WN624 and low-pressure-evolved strain WN1106 at 50 mbar were identified by microarray analysis. Among these was a cluster of three candidate genes (des, desK, and desR), whose mRNA levels in WN1106 were higher than the ancestor on the microarrays. Up-regulation of these genes was confirmed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis. The des, desK, and desR genes encode the Des membrane fatty acid (FA) desaturase, the DesK sensor kinase, and the DesR response regulator, respectively, which function to maintain membrane fluidity in acute response to temperature downshift. Pressure downshift caused an up-regulation of des mRNA levels only in WN1106, but expression of a des-lacZ transcriptional fusion was unaffected, which suggests that des regulation was different in response to temperature versus pressure downshift. Competition experiments showed that inactivation of the des gene caused a slight, but statistically significant, loss of fitness of strain WN1106 at 50 mbar. Further, analysis of membrane FA composition of cells grown at 1013 versus 50 mbar revealed a decrease in the ratio of unsaturated to saturated FAs but an increase in the ratio of anteiso- to iso-FAs. The present study represents a first step toward identification of molecular mechanisms by which B. subtilis could sense and respond to the novel environmental stress of low pressure.

Effect of Solar Radiation on Severity of Soybean Rust

Soybean rust (SBR), caused by Phakopsora pachyrhizi, is a damaging fungal disease of soybean (Glycine max). Although solar radiation can reduce SBR urediniospore survival, limited information is available on how solar radiation affects SBR progress within soybean canopies. Such information can aid in developing accurate SBR prediction models. To manipulate light penetration into soybean canopies, structures of shade cloth attenuating 30, 40, and 60% sunlight were constructed over soybean plots. In each plot, weekly evaluations of severity in lower, middle, and upper canopies, and daily temperature and relative humidity were recorded. Final plant height and leaf area index were also recorded for each plot. The correlation between amount of epicuticular wax and susceptibility of leaves in the lower, middle, and upper canopies was assessed with a detached leaf assay. Final disease severity was 46 to 150% greater in the lower canopy of all plots and in the middle canopy of 40 and 60% shaded plots. While daytime temperature within the canopy of nonshaded soybean was greater than shaded soybean by 2 to 3°C, temperatures recorded throughout typical evenings and mornings of the growing season in all treatments were within the range (10 to 28.5°C) for SBR development as was relative humidity. This indicates temperature and relative humidity were not limiting factors in this experiment. Epicuticular wax and disease severity in detached leaf assays from the upper canopy had significant negative correlation (P = 0.009, R = -0.84) regardless of shade treatment. In laboratory experiments, increasing simulated total solar radiation (UVA, UVB, and PAR) from 0.15 to 11.66 MJ m(-2) increased mortality of urediniospores from 2 to 91%. Variability in disease development across canopy heights in early planted soybean may be attributed to the effects of solar radiation not only on urediniospore viability, but also on plant height, leaf area index, and epicuticular wax, which influence disease development of SBR. These results provide an understanding of the effect solar radiation has on the progression of SBR within the soybean canopy.

Genetic differentiation in natural populations of a Keystone Bunchgrass (Aristida stricta) across its native range

Aristida stricta Michx. (Poaceae) is a perennial bunchgrass native to the Southeastern Coastal Plain of North America where it is a keystone species in the longleaf pine savannas and slash pine flatwoods from southeastern North Carolina to Florida, and westward to the coast of Mississippi. We examined genetic relationships within and among ten populations of A. stricta by using eight inter-simple sequence repeat (ISSR) markers to generate band frequency data for 32 individuals from each sampled population. An analysis of molecular variance showed that 38% of the variation resided among populations while 62% was attributable to variation within populations. Grouping the populations by habitat or by geographic location did not show significant differentiation between the groups. Overall, pair-wise geographic and genetic distances were not correlated. Data indicate that while individuals within each population are genetically diverse, there seemingly are barriers to gene flow across populations leading to their divergence. Each population contains several exclusive loci suggesting that limited gene flow and/or genetic drift are likely leading to this pattern of localization. Our results, coupled with those of the previous studies that presented evidence for local adaptation and phenotypic differences among populations, suggest that there is sufficient differentiation among populations of this species to warrant: (1) maintenance of the existing genetic diversity at individual sites, and (2) use of local seed and plant sources for conservation projects.

First Report of Turnip mosaic virus Infecting Brassica carinata (Ethiopian Mustard) in the United States

Brassica carinata L. Braun (Ethiopian mustard) is an annual oil seed crop currently being evaluated for its potential use as a source of biofuel. Due to its high content of erucic acid, it provides a biodegradable non-fossil fuel feedstock that has many applications ranging from biofuels to other industrial uses such as polymers, waxes, and surfactants. Moreover, high glucosinolate content adds the scope of B. carinata being used as a bio-fumigant. B. carinata is amenable to low input agriculture and has great economic potential to be used as a winter crop, especially in the southeastern United States. Virus-like leaf symptoms including mosaic, ringspot, mottling, and puckering were observed on B. carinata (cvs. 080814 EM and 080880 EM) in field trials at Quincy, FL, during spring 2013, with disease incidence of >80%. A more extensive survey of the same field location indicated that mosaic symptoms were the most common. Viral inclusion assays (1) of leaves with a range of symptoms indicated the presence of potyvirus-like inclusion bodies. Total RNA extracts (RNeasy Plant Mini Kit, Qiagen Inc., Valencia, CA) from six symptomatic samples and one non-symptomatic B. carinata sample were subjected to reverse transcription (RT)-PCR assays using SuperScript III One-Step RT-PCR System (Invitrogen, Life Technologies, NY), and two sets of potyvirus-specific degenerate primers MJ1-F and MJ2-R (2) and NIb2F and NIb3R (3), targeting the core region of the CP and NIb, respectively. The RT-PCR assays using the CP and NIb specific primers produced amplicons of 327 bp and 350 bp, respectively, only in the symptomatic leaf samples. The obtained amplicons were gel-eluted and sequenced directly (GenBank Accession Nos. KC899803 to KC899808 for CP and KC899809 to KC899813 for NIb). BLAST analysis of these sequences revealed that they came from Turnip mosaic virus (TuMV). Pairwise comparisons of the CP (327 bp) and NIb (350 bp) segments revealed 98 to 99% and 96 to 98% nucleotide identities, respectively, with corresponding sequences of TuMV isolates. These results revealed the association of TuMV with symptomatic B. carinata leaf samples. Although TuMV has been reported from B. carinata in Zambia (4), this is the first report of its occurrence on B. carinata in the United States. Considering the importance of B. carinata as a biofuel source, this report underscores the need for developing effective virus management strategies for the crop. References: (1) R. G. Christie and J. R. Edwardson. Plant Dis. 70:273, 1986. (2) M. Grisoni et al. Plant Pathol. 55:523, 2006. (3) L. Zheng et al. Plant Pathol. 59:211, 2009. (4) D. S. Mingochi and A. Jensen. Acta Hortic. 218:289, 1988.

Cattle Grazing Affects Cotton Root Dimensions and Yield in a Bahiagrass-based Crop Rotation

With increasing production costs in the southeastern US, cropping systems that improve agronomic and economic sustainability are critical. A sod-based system utilizing a two year rotation of establishing and grazing bahiagrass (Paspalum notatum Fluegge) followed by a year of peanut (Arachis hypogaea L.) and a year of cotton (Gossypium hirsutum L.) was established in Marianna, FL. The effect of grazing in this system on root dimensions (length, diameter, and surface area) of cotton was monitored using a mini-rhizotron technique. There were differences in several root parameters between grazed and non-grazed plots (256 m²) including: cotton root length (1260 mm grazed vs. 844 mm non-grazed), surface area (1727 mm² grazed vs. 1105 mm² non-grazed), and diameter (3.09 mm grazed vs. 2.51 mm non-grazed). The roots in the 15 to 30-cm-depth profile had greater length and surface area in the grazed plots with the greatest root surface areas in the deepest soil profile (75-90 cm). Grazed plots also exceeded nongrazed plots in seed cotton yield (3967 kg/ha vs. 3335 kg/ha) and lint (1672 kg/ha vs. 1444 kg/ha). Increases in root architecture associated with cattle grazing in this system may enable cotton to better withstand adverse environmental conditions such as drought stress, thereby improving long-term sustainability. Utility of Sod-based Systems in the Southeastern US The number of farms in the southeastern US is declining due to consolidation into larger operations and a shift to other uses like pine production and other forestry-based applications (20). This situation makes it necessary to develop robust cropping systems that are profitable while conserving the environment through sustainable agricultural practices. In order to achieve these goals, yields obtained from sustainable systems must be increased while maintaining or even decreasing production costs. This will enable small farms of less than 200 ha to remain competitive in national and international markets (37). In the panhandle of Florida and much of the southeast US, the most common crop rotation sequence is peanut (Arachis hypogaea L.), followed by two years of cotton (Gossypium hirsutum L.). While both cotton and peanut are major crops in the US, both are marginally profitable due to high production costs and uncontrollable weather events that reduce yields. One technology that can often reduce costs associated with fuel and irrigation is conservation tillage; currently nearly 35% of the total US cotton area implements some form of reduced tillage (6). An essential element to most conservation tillage systems is the use of cover crops during the winter period prior to the summer crop to reduce erosional soil losses, increase water holding capacity, and decrease evaporative losses during the cropping season.

Cattle grazing affects peanut root characteristics in a bahiagrass-based crop rotation system

ABSTRACT The critical aspect of production agriculture in the southeastern US with increasing associated costs is to improve economic and agronomic sustainability. A four yr sod-based rotation system consisting of two yr of bahiagrass (Paspalum notatum Fluegge) (grazed or non-grazed) followed by a yr of peanut (Arachis hypogaea L.) and a yr of cotton (Gossypium hirsutum L.), each with winter cover crop (grazed or non-grazed) was established in Marianna, FL. The effect of grazing on root parameters (length, volume, surface area, and diameter) of peanut was observed using a mini-rhizotron. There were differences in several root parameters between grazed and non-grazed plots including: peanut root length (307 mm in grazed vs 167 mm in non-grazed), volume (50 mm3 in grazed vs. 23 mm3 in non-grazed), surface area (399 mm2 in grazed vs. 197 mm2 in non-grazed), and diameter (2.4 mm in grazed vs. 1.7 mm in non-grazed). Roots at the 45-60 cm and 60-75 cm depths had significantly greater length in the grazed than t...