{"id":263,"date":"2016-08-20T08:44:37","date_gmt":"2016-08-20T08:44:37","guid":{"rendered":"http:\/\/www.labnetwest.colourfi.com\/?p=263"},"modified":"2025-12-22T06:03:51","modified_gmt":"2025-12-21T22:03:51","slug":"plant-tissue-culture","status":"publish","type":"post","link":"https:\/\/ciderhousetest.com\/labnetwest26\/plant-tissue-culture\/","title":{"rendered":"Plant Tissue Culture:"},"content":{"rendered":"

FROM THE CSIRO LAB AT SCITECH:<\/strong>
\nThere is a program in the CSIRO Lab at Scitech called PLANTECH.
\nAnother sterile technique of tissue culture, carried out in laminar flow cabinets, is being offered for high schools, called an embryo rescue. It is also used in conservation biotechnology at Kings Park, plus a huge range of other plant biotechnology and botanic activities.
\nAgar is not actually needed to do enquiries into the effect of plant hormones on growth.
\nPlant rooting hormones and potting mix or vermiculite can simply be used. The best way to measure roots is to harvest the plant, wash it out and use a grid that you lay the roots across and count the number of intersections of roots across the lines.
\nHormones can be bought at gardening and hardware stores and probably at scientific suppliers.
\nAGAR CULTURE:<\/strong>
\nLow grade agar powder is fine.
\nSugar needs to be in the agar, plus a complete fertiliser and M&S (Murashige and Skoog) basal medium
\nSugar encourages fungal and bacterial growth, so the vials must be sterilised in an autoclave or pressure cooker. Be aware that some types melt, some don\u2019t.
\nPLANT TISSUE CULTURE:<\/strong>
\nPrepare media for agar vial tissue culture at least one day before class
\nMix in 2 litre beaker:
\n1 bottle MS (nutrients) 4 g (rinse out)
\n1 litre distilled or tap water
\n30 g sucrose. Sucrose aids photosynthesis but also promotes microbial growth
\nAdjust pH to 6 with sodium hydroxide solution or HCl (10%), added drop-wise with a pipette
\n8 g agar
\nLet it boil to mix agar (in microwave), approximately 5 minutes
\nPour into polycarbonate vials ready to autoclave (sterilise)
\nEg: 10 ml in 120 mL vial. Smaller containers mean that can get more out of the 1 litre agar.
\nPrepare tools for autoclave or pressure cooker.
\nRequirements:
\nLivingstone Mediplast-pressure tape (autoclave masking tape with colour change)
\n2 forceps, 2 scalpels or dissecting scissors
\nSterilise laminar flow cabinets
\nWipe down regularly with ethanol
\nLeave UV light on for at least 20 minutes
\nOr do this in a sterilised aquarium. This is not as good but much cheaper.
\nCLONING:<\/strong>
\nYou can propagate plants from softwood cuttings.
\nAuxins (plant hormones) induce new root formation.
\nIn horticulture, auxins, especially Indole-3-butyric acid and Indole-3-acetic acid, are commonly applied to stimulate root growth when taking cuttings of plants.
\nIn a couple of weeks, plants will have adventitious roots.
\nAdventitious roots differ from the more usual root formation of branches of a main primary root, and instead originate from the stem, branches, leaves, or old woody roots.
\nLOOKING AFTER YOUR CLONED PLANT:<\/strong>
\nThe cloned plant can be transplanted within three weeks, when it has started to develop adventitious roots. It can transplanted it into a pot with potting mix.
\nThe new plant is a clone as it has the same genetic material (DNA) as the parent plant from which it was cut.
\nPLANT TISSUE CLONING:<\/strong>
\nWhat to do
\n1. Wash and dry a pair of scissors. Clean scissors are required to avoid infection and disease.
\n2. On the petri dish, cut a piece of shoot from a main stem of your parent plant making sure there is a node.
\n3. Remove most of the older leaves so that the plant doesn\u2019t use too much water or die because of a lack of roots. Place unwanted plant material in the bin.
\n4. Keep only the young growing shoot. This called an explant.
\n5. Make sure that your explant is the right way up.
\n6. Dip the bottom end of the explant into the auxin (plant hormone).
\n7. Shake off any excess root hormone.
\n8. Remove the lid from your vial.
\n9. Push the explant into the agar.
\n10. Replace the lid.
\nIt is important to shake off surplus auxin (plant hormone) as too much auxin will stop root growth. A small amount of root hormone will make adventitious roots grow.<\/p>\n","protected":false},"excerpt":{"rendered":"

FROM THE CSIRO LAB AT SCITECH: There is a program in the CSIRO Lab at Scitech called PLANTECH. Another sterile technique of tissue culture, carried out in laminar flow cabinets, is being offered for high schools, called an embryo rescue. It is also used in conservation biotechnology at Kings Park, plus a huge range of […]<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[4],"tags":[23,24],"class_list":["post-263","post","type-post","status-publish","format-standard","hentry","category-experiments","tag-cloning","tag-plants"],"_links":{"self":[{"href":"https:\/\/ciderhousetest.com\/labnetwest26\/wp-json\/wp\/v2\/posts\/263","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/ciderhousetest.com\/labnetwest26\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/ciderhousetest.com\/labnetwest26\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/ciderhousetest.com\/labnetwest26\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/ciderhousetest.com\/labnetwest26\/wp-json\/wp\/v2\/comments?post=263"}],"version-history":[{"count":0,"href":"https:\/\/ciderhousetest.com\/labnetwest26\/wp-json\/wp\/v2\/posts\/263\/revisions"}],"wp:attachment":[{"href":"https:\/\/ciderhousetest.com\/labnetwest26\/wp-json\/wp\/v2\/media?parent=263"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/ciderhousetest.com\/labnetwest26\/wp-json\/wp\/v2\/categories?post=263"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/ciderhousetest.com\/labnetwest26\/wp-json\/wp\/v2\/tags?post=263"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}