Sufficient yields of high-quality RNA are needed for next-generation sequencing and high-throughput real-time polymerase chain reaction analyses. In the case of strawberry (Fragaria ×ananassa) fruits, successful RNA isolation requires removal of abundant inhibitory substances (polysaccharides and polyphenols) that greatly reduce quality and yield. In this study, we applied various combinations of RNA isolation protocols directed at reproductive organs. The best manual isolation method involved nonionic polymer and modified acid guanidinium thiocyanate-phenol-chloroform treatments followed by phenol/chloroform/isoamyl alcohol extraction. Compared with other methods, this approach gave significantly higher yields [84.0 µg/g fresh weight (FW)] of RNA of greater purity (A260/A280 = 1.99; A260/230 = 1.51). Better-quality RNA (A260/230 = 2.11) was obtained using an automated method, but the yield was lower (18.1 µg/g FW) than that obtained manually. This automated method consisted of pretreatment with nonionic polymer followed by a silica-based system extraction. Although RNA of sufficient quality [RNA Integrity Number (RIN) ≥ 6.5 and 28S/18S ≥ 1.0] for RNA sequencing was obtained from receptacles using both automated and manual methods, the manual method yielded high-quality RNA from achenes and anthers. The automatic method features 6-fold faster high-throughput capacity, whereas the manual method has wider applicability to different tissues.
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Misaki Ishibashi, Takeshi Nabe, Yoko Nitta and Yuichi Uno
Hardeep Singh, Bruce Dunn, Mark Payton and Lynn Brandenberger
Nutrient-film technique (NFT) trials were conducted to quantify the effect of two different water-soluble hydroponic fertilizers (5N–4.8P–21.6K and 5N–5.2P–21.6K) on different cultivars of lettuce (Lactuca sativa), basil (Ocimum basilicum), and swiss chard (Beta vulgaris). Results indicated swiss chard yield was affected only by cultivars, with Fordhook Giant producing the greatest fresh weight across fertilizer treatments. For lettuce production, interaction between fertilizers and cultivars was significant. ‘Mirlo’ and ‘Rubysky’ had greater growth compared with other cultivars in both fertilizers, whereas Dragoon performed well using 5N–4.8P–21.6K, but not 5N–5.2P–21.6K. For basil, dry weight production showed a significant interaction between fertilizers and cultivars. ‘Largeleaf’ produced greater dry weight with 5N–4.8P–21.6K, whereas ‘Lemon’ produced greater dry weight with 5N–5.2P–21.6K. For nutrient concentration of leaves, the concentrations were within the recommended range for lettuce when fertilized with 5N–5.2P–21.6K. Nutrient concentrations varied by nutrient from the recommended range for basil, but there was no significant difference between fertilizers. For swiss chard, the nutrient concentrations were in the recommended range and there was no difference between fertilizers. Therefore, growers may need to use more than one type of fertilizer for different lettuce and basil cultivars for optimum production, whereas swiss chard cultivars can be selected based on yield regardless of fertilizer.
Brian A. Mitchell, Mark E. Uchanski and Adriane Elliott
This research assessed fruit load management and production techniques for cultivating indeterminate tomatoes in a high tunnel under intensive organic management. The successful production of high-quality, high-yielding crops is important for fruit and vegetable producers, especially growers using high tunnels. High tunnels are well-suited to organic farming and can be used to grow many valuable specialty crops. Fruit load management is practiced in fruit production (e.g., apples, peaches, and grapes), but there is lack of consensus concerning the effectiveness of fruit cluster pruning on tomato (Solanum lycopersicum) and its impact on fruit yield, quality, and marketability. In addition, there is no published research on tomato cluster pruning in certified organic systems or intensively managed high tunnels (e.g., densely planted, trellised, vegetatively pruned plants) for the Front Range of Colorado. In 2016 and 2017, a randomized complete block design was used to test the effects of cluster pruning within a high tunnel on certified organic land at Colorado State University’s (CSU’s) Agricultural Research, Development, and Education Center, South. Two treatments and three tomato cultivars were selected for the study; the treatment–cultivar combinations were replicated six times within a high tunnel. The treatments involved reducing fruit loads to three fruit and six fruit per cluster, whereas plants with unpruned clusters, which naturally developed as many as 10 fruit, served as the control. Tomato cultivars evaluated were ‘Cherokee Purple’, a widely studied heirloom, and two hybrids: ‘Jet Star’ and ‘Lola’. Parameters measured included total yield, individual fresh fruit weight, soluble solids content (SSC), marketable yield, and nonmarketable yield. Individual fresh fruit weight increased for both hybrids in the three-fruit treatment, averaged over two growing seasons. ‘Cherokee Purple’ did not respond to the cluster pruning treatments. There was no decrease in total yield, across all cultivars, between treatments and the unpruned control. However, ‘Jet Star’ yielded more than the other two cultivars. In addition, SSC and marketability measurements were more influenced by cultivar than cluster pruning treatments. ‘Lola’ had a significantly greater SSC than the other two cultivars. ‘Jet Star’ had the greatest marketable yields of all cultivars tested whereas ‘Cherokee Purple’ produced greater nonmarketable (cull) yields. Cluster pruning produced larger organic tomatoes without reducing yield or quality for two of the three cultivars used in the study. Cultivar selection remains one of the greatest factors in determining yield, quality, and marketability of a crop.
Lyn A. Gettys and Kimberly A. Moore
Wetland restoration is critical for improving ecosystem services, but many aquatic plant nurseries do not have facilities like those typically used for large-scale plant production. We questioned if we could grow littoral aquatic plant species in a variety of substrates and irrigation methods similar to those used for traditional greenhouse production. Plants were grown in pots with drainage holes that were filled with potting substrate, topsoil, coarse builders’ sand, or a 50/50 mix of topsoil and builders’ sand. These substrates were amended with 2 g of 15N–3.9P–10K controlled-release fertilizer per liter of substrate and were watered using either overhead irrigation or subirrigation. Plants were grown for 16 weeks, then scored for quality and height before a destructive harvest. Blue-eyed grass (Sisyrinchium angustifolium) and arrow arum (Peltandra virginica) performed best when subirrigated and cultured in potting substrate or sand. Golden club (Orontium aquaticum) and lemon bacopa (Bacopa caroliniana) grew best when plants were cultured in potting substrate and maintained under subirrigation. These experiments provide a framework for using existing greenhouses to produce these littoral species and give guidance to growers who wish to produce plants for the restoration market.
Production and use of sweet olive (Osmanthus armatus), fragrant tea olive (O. fragrans), holly tea olive (O. heterophyllus), and fortune’s osmanthus (O. xfortunei) as a landscape plant is currently limited to U.S. Department of Agriculture (USDA) Hardiness Zones 7 to 10, and nursery growers wish to extend the range of these species into colder climates. To provide recommendations to growers and landscapers and inform breeding efforts for cold-hardiness improvement, a replicated trial was conducted in a USDA Hardiness Zone 6b/7a transition zone. Fifteen cultivars and two unnamed accessions representing four species were evaluated for growth, stem necrosis, and flowering in a pot-in-pot production system from 2015 to 2017. One-half of the plants in each cultivar were moved to winter protection each November and returned to the field each May. There were significant differences in growth and cold-hardiness among cultivars. Percent increase in the growth index after three growing seasons for winter-exposed accessions of sweet olive, fortune’s osmanthus, fragrant tea olive, and holly tea olive averaged 867%, 1175%, 155%, and 6361%, respectively. Percent stem necrosis in May 2017 for sweet olive, fortune’s osmanthus, fragrant tea olive, and holly tea olive averaged 1.1%, 2.7%, 44.8%, and 20.2%, respectively. The most cold-tolerant accessions based on stem necrosis and growth index of winter-exposed plants were ‘Kaori Hime’, ‘Hariyama’, ‘Shien’, ‘Head-Lee Fastigate’, and ‘Rotundifulius’ holly tea olive, ‘San Jose’ fortune’s osmanthus, and ‘Longwood’ sweet olive. Of these cultivars, Kaori Hime, San Jose, and Longwood flowered under winter-exposed conditions. All fragrant tea olive cultivars were damaged by winter exposure. ‘Fodingzhu’ was the only fragrant tea olive cultivar that flowered each year under winter-exposed conditions. Evaluation and breeding efforts are continuing to extend the range for production and growth of this genus.
Chad E. Finn, Bernadine C. Strik, Brian M. Yorgey, Mary E. Peterson, Patrick A. Jones, Jungmin Lee, Nahla V. Bassil and Robert R. Martin
Xuan Wu, Shuyin Liang and David H. Byrne
Plant architecture is a crucial trait in plant breeding because it is linked to crop yield. For ornamental crops such as roses, plant architecture is key for their aesthetic and economic value. In 2015, six rose plant architectural traits were evaluated on 2- to 3-year-old plants of F1 rose populations in May and December in College Station, TX, to estimate variability and heritability. The traits included plant height, the number of primary shoots, the length of primary shoots, the number of nodes on the primary shoot, the number of secondary shoots per primary shoot, and the number of tertiary shoots per primary shoot. Among these traits, plant height, the number of primary shoots, and the length of primary shoots showed a substantial amount of variability, whereas the number of secondary and tertiary shoots per primary shoot were skewed toward zero. Using a random effects model restricted maximum likelihood (REML) analysis, the architectural traits demonstrated low to moderate narrow-sense heritability (0.12–0.50) and low to high broad-sense heritability (0.34–0.92). Plant height and the number of primary shoots changed little after the first growth phase, whereas the other four traits were affected greatly by the genotype-by-environment (growth phase) interaction.
M. Taylor Perkins, Anna Claire Robinson, Martin L. Cipollini and J. Hill Craddock
Phytophthora cinnamomi Rands, the causal pathogen of phytophthora root rot (PRR) of chestnut, is one of the main obstacles to growth of american chestnut [Castanea dentata (Marsh.) Bork.] in the southern part of its distribution. To facilitate introgression of PRR resistance of chinese chestnut (C. mollissima Blume) into a C. dentata genetic background, we assessed the disease resistance of 10 interspecific hybrid families derived from potentially resistant C. mollissima cultivars. Hybrid progeny were inoculated with P. cinnamomi in the nursery and assessed for root lesion severity after 1 year of growth. Asymptomatic plants were transplanted to a P. cinnamomi-positive orchard and evaluated for survival midway through the following growing season. During the nursery experiment, 8 of 10 hybrid families were not significantly different from susceptible C. dentata controls for average disease resistance scores. However, multiple asymptomatic individuals were identified in each of the eight families. Two of the 10 hybrid families were not significantly different from the resistant C. mollissima and C. henryi controls. In the P. cinnamomi-positive orchard, the prescreened hybrid families displayed a greater proportion of survivors than backcross families that had not been prescreened for P. cinnamomi resistance. Hybrid plants that have survived 2 years of growth in P. cinnamomi-infested potting media and soils represent an important step toward the production of genetically diverse chestnut populations in the southeastern United States that combine the PRR resistance of C. mollissima with the morphology and local adaptation of C. dentata.
Robert Andrew Kerr, Lambert B. McCarty, Philip J. Brown, James Harris and J. Scott McElroy
Summer annual grassy weeds such as goosegrass (Eleusine indica L. Gaertn.) continue to be problematic to control selectively with postemergence (POST) herbicides within turfgrass stands. In recent years, reduced performance by certain herbicides (e.g., foramsulfuron), cancellation of goosegrass-specific herbicides (e.g., diclofop-methyl), and cancellation and/or severe use reductions of other herbicides [e.g., monosodium methanearsonate (MSMA)] have limited the options for satisfactory control and maintenance of an acceptable (≤30% visual turfgrass injury) turfgrass quality. Currently available herbicides (e.g., topramezone and metribuzin) with goosegrass activity typically injure warm-season turfgrass species. The objectives of this research were to evaluate both ‘Tifway 419’ bermudagrass [Cynodon dactylon (L.) Pers. ×Cynodon transvaalensis Burtt-Davy] injury after treatment with POST herbicides, and to determine whether irrigating immediately after application reduces turfgrass injury. Treatments were control (± irrigation); topramezone (Pylex 2.8C; ± irrigation); carfentrazone + 2,4-D + dicamba + 2-(2-methyl-4-chlorophenoxy) propionic acid (MCPP) (Speedzone 2.2L; ± irrigation); carfentrazone + 2,4-D + dicamba + MCPP in combination with topramezone (± irrigation); metribuzin (Sencor 75DF; ± irrigation); mesotrione (Tenacity 4L; ± irrigation); simazine 4L (±irrigation); and mesotrione + simazine (± irrigation). Irrigated treatments were applied immediately with a hand hose precalibrated to apply 0.6 cm or 0.25 inch (≈6.3 L). Visual turfgrass injury for combined herbicide treatments for the irrigated plots was 6% 4 days after treatment (DAT), 12% 1 week after treatment (WAT), 17% 2 WAT, and 6% 4 WAT, whereas nonirrigated plots had turfgrass injury of 14% at 4 DAT, 31% 1 WAT, 35% 2 WAT, and 12% 4 WAT. Irrigated pots had normalized differences vegetative indices (NDVI) ratings of 0.769 at 4 DAT, 0.644 at 1 WAT, 0.612 at 2 WAT, and 0.621 at 4 WAT, whereas nonirrigated plots had the lowest (least green) turfgrass NDVI ratings of 0.734 at 4 DAT, 0.599 at 1 WAT, 0.528 at 2 WAT, and 0.596 at 4 WAT. These experiments suggest turfgrass injury could be alleviated by immediately incorporating herbicides through irrigation.
Suzanne Stone, George Boyhan and Cecilia McGregor
Watermelon [Citrullus lanatus (Thunb.) Matsum. & Nakai] cultivars exhibit diverse phenotypic traits, yet are derived from a narrow genetic base. Heirloom cultivars, and to a lesser extent modern open-pollinated (OP) cultivars, are perceived to contain vital genetic variation that is critical for biodiversity conservation and crop improvement. The objective of this study was to characterize the diversity of six heirloom and open-pollinated watermelon cultivars that are popular among U.S. organic, direct-market, and home gardeners. An additional evaluation was conducted to determine whether significant phenotypic and genotypic variation existed among seed lots sourced from different commercial seed vendors. Important horticultural traits such as days to germination, days to first flower, yield, and fruit quality were measured over two field seasons. Genetic diversity was estimated using 32 simple sequence repeat (SSR) markers. Significant differences in horticultural traits among seed lots in both years were observed only in days to germination and first male flower, which may be a consequence of vendor differences in seed storage and quality control. Heirloom ‘Moon and Stars’ and modern OP ‘Sugar Baby’ were the most genetically distinct from the other cultivars and heirloom ‘Georgia Rattlesnake’ was determined to be highly related to the modern OP ‘Charleston Gray’. The two heirloom cultivars were observed to have lower average gene diversity than the modern cultivars. Heirloom ‘Moon and Stars’ contained significant genetic variation among seed lots, yet heirloom ‘Georgia Rattlesnake’ contained none. These findings suggest that genetic variation can be more accurately attributed to pedigree and foundation seed maintenance practices than to the “heirloom” designation per se. The variation reported in this study can be used to inform conservation and breeding efforts.