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Propagating Cynthiana/Norton Grapes Basic Overview and MTSU Research Update Tony V. Johnston, PhD, CFS, MS, MPH Professor School of Agribusiness and Agriscience Middle Tennessee State University Overview • Propagation Basics • Dormant (Hardwood) Propagation • Softwood Tip Propagation • Bench Grafting • Tissue Culture • From the Greenhouse to the Vineyard • Hardwood Propagation Issues • Propagate or Not? • MTSU Research Updates • Endophytes and their Impacts • Current and Future MTSU Research Goals • Research Credits • Questions Propagation Basics • Always use clean, sharp knife and sterile rooting medium (no fertilizer) • Never use pruning shears or knife used to cut other plants without prior sterilization – Can transfer infectious viruses, mold, etc. • High humidity environment is essential (no roots!) • 75oF minimum temp at all times • Good light– use a grow light if necessary • • • • • • Dormant (Hardwood) Propagation Traditional method Pencil diameter, 1-1.5 ft long Must be viable wood (green pith) Collect after vines are completely dormant Remove all shoots and leaves Set in sand, vermiculite/soil, or potting soil – Largest diameter end down; dip in rooting hormone (e.g. Roottone® or Hormonex®) • Constant temperature above 60oF Softwood Tip Propagation – Collect cuttings anytime during growing season – Cut 3-4 inches from growing tip end – Strip all but 3-4 leaves on growing end of shoot – Dip end in rooting hormone (e.g. Roottone® or Hormonex®) – Place in sterile sand, vermiculite or peat moss – Keep moist but not soaking – Minimum temp 72oF; warmer better Bench Grafting (pt. 1) • Dormant propagation technique • Not widely employed at home • Essential for protection of non-phylloxera resistant varieties of grapes • Must have rootstock and scion • Both rootstock and scion must be very similar in diameter Bench Grafting (pt. 2) • Both rootstock and scion are cut in identical ways (slant or z) • With machine, omega cut can be made • Join at cut, aligning carefully • Wrap union with grafting tape and allow to heal • Transfer to ground after viability is established • Do not remove grafting tape! Tissue Culture (pt 1) • Definitely not a home-based technique • Sterile environment and technique essential • Plant cells are extracted from carefully washed and sterilized plant parts (leaf, stem or root) • Place cells on specialized growth medium • Cells grow into undifferentiated (nonspecialized) “callus” Tissue Culture (pt. 2) – Callus is then transferred to different medium and conditions adjusted to stimulate the development of roots and stems – Hormone and medium composition is different for almost all plants – End result is a true clone– an exact replica of the source cells Greenhouse/Lab to Vineyard • Propagated vines should be planted outdoors ASAP • Vines don’t do well in a greenhouse environment • Best to plant in spring after last frost • Two to three years to first harvest Cynthiana/Norton Hardwood Propagation Issue • 20 – 40% success rates typical – Success = viable plant • Common result: leaf production without root development • Must compensate by trying to root hundreds to get tens of successes – Time and resource intensive with low return on investment Propagate or Not? • Commercial plants are available to purchase • Best to purchase from nurseries “close” to Tennessee • If you have a “facility” in which to grow out your cuttings, home propagation is inexpensive, albeit time consuming and inefficient – Can be significantly more cost effective than purchasing vines from a nursery MTSU Research Update • Focused on finding ways to better propagate Cynthiana/Norton grapes • Both hardwood and tissue culture techniques being studied • A follow-on to research conducted in Honduras – Enormous difficulty in rooting dormant vines – Complete failure of tissue culture efforts due to mold growth on plates MTSU Hardwood Propagation Research • Evaluating: – Timing of hardwood collection (12 month project) – Efficacy of rooting hormone use – Timing of planting of hardwood propagated plants in the vineyard • Is dogma correct? MTSU Tissue Culture Research • Initial attempts to culture Cynthiana/Norton grapes totally unsuccessful • 100% failure to stimulate mold-free callus formation • While using different antifungal compounds and trying different presampling techniques discovered an endophyte in the vines What is an Endophyte? • Organisms, often fungi and bacteria, that live between living plant cells • Relationship with the plant varies from symbiotic to bordering on pathogenic • Extremely unusual and valuable organic substances are produced that may assist in helping solve not only human health, but plant and animal health problems – From http://plantsciences.montana.edu/facultyorstaff/faculty/strobel/endophytes.html Endophyte Impact • Being a mold and living between the cells, it always contaminates tissue culture medium • Must eliminate endophyte to allow for tissue culture • Role of endophyte in Cynthiana/Norton grapes unclear • May play a role in the difficulty to propagate via hardwood culturing Current MTSU Research Outcomes and Focus • MTSU research has resulted in technique to eliminate endophyte without killing plant tissue • Now researching techniques to take endophyte free tissue to plantlet development • Looking to create line of endophyte free plants Future MTSU Research • Focus on – Exploration of role of endophyte in Cynthiana/Norton grapevines – Determine whether endophyte-free Cynthiana/Norton are viable and can remain endophyte-free once planted – Determine productivity of potentially endophyte-free Cynthiana/Norton – Determine quality of fruit produced by potentially endophyte-free Cynthiana/Norton – Identify Cynthiana/Norton endophyte Research Credits • Dr. John DuBois, MTSU Biology Professor • Mr. Shannon Smith, MTSU Biology Graduate Student • Ms. Amanda Leachman, MTSU Biology Undergraduate Student • Ms. Aimee Wilson, MTSU Biology Undergraduate Student Questions?