Massachusetts Agricultural Experiment Station

In vitro maturation, in vitro fertilization, and embryo development will be according to procedures widely published in the literature and frequently used in the laboratory. In vitro maturation and fertilization will be conducted in 400 μl drops at 38.5°C and 5% CO2 in a humidified atmosphere, while embryo culture will be carried out in 100 μl drops under mineral oil of KSOM– 0.4% BSA medium (EmbryoMax, Chemicom International, USA) at 38.5°C and 5% CO2, 5% O2 and 90% N2, in a humidified atmosphere.

This research will help to understand and develop practices that will improve soil fertility by enhancing the efficiency of use of nutrients supplied by organic or conventional fertilizers. The increased efficiency may help improve yields and quality of plant-derived foods and allow for production of nutritjous, healthful foods for consumers. The accumulation of elemental nutrients in these foods is expected to be enhanced by biochar compounded with conventional or organic fertilizers.

Modern cranberry producers utilize a diverse array of pest management and horticultural techniques to produce a marketable crop. In a typical commercial setting, three to five fungicide applications are made during the growing season and resultant field rot levels range from less than 1% to 15%. Fruit rot infected lots beyond 12% are heavily discounted and lots with >20% are not accepted by cranberry handlers (cranberry processing industry). Some growers are seeing their entire delivery of fruit rejected for rot levels exceeding the maximum allowed.

Methodology

The CRISPR/Cas9 technology is increasingly popular in plant research to generate mutants in target genes. In CRISPR/Cas9, the gRNA mediates a double-stranded DNA break, which when repaired often leaves small mutations. These mutations can knock out the function of the target gene, but only when all alleles of the gene are mutated. Because the repair is imprecise, the exact mutations will differ between cells.

The study area is the Connecticut River watershed, extending from the U.S.-Canada border north to the Long Island Sound south. The watershed drains over 28,490 Km2 and contains 390 towns and cities with a population of approximately 2.3 million. The watershed has 79% forest cover and 11% agriculture (Marshall and Randhir, 2008b). Modeling the dynamics of FEW systems requires using multiple spatial and temporal processes of watershed hydrology, built energy, and agriculture subsystems.

 We will employ two well-characterized and widely used in vitro cell culture systems to address each of the objectives proposed. Namely, 3T3-L1 preadipocytes and RAW264.7 macrophages (available in both PI’s lab). Initial cell culture experiments will test the primary hypothesis that treatment of cranberry extract (CBE) promotes fatty acid oxidation and mitochondrial thermogenic biogenesis in 3T3-L1 adipocytes and anti-inflammatory effects in RAW264.7 macrophages.

Tree growth, post-establishment [defined as a resumption of pre-transplant growth rate (Struve and Joly 1992)], will be determined by measuring caliper of the established 48 research oak (Q. bicolor, Q. rubra) trees, annually for the next four years (preferably longer, but contingent upon funding). Annual increment will be analyzed using piecewise regression.

1) Program Participants: food delivery through Senior Food Boxes, Surveys to assess food security, food access, healthy food consumption, and program perceptions and experiences; 2) Program Staff/Producers: Interviews with key
informants (i.e., program staff and producers/farmers) to assess program efficiencies, program challenges, and ideas for addressing those challenges.