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\author{woozing}
\pdfinfo{
  /Title (ptc-c4-explant-sterilization.pdf)
  /Creator (Cheatography)
  /Author (woozing)
  /Subject (PTC - C4 (Explant Sterilization) Cheat Sheet)
}

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        \hspace*{-6pt}\includegraphics[width=5.8cm]{/web/www.cheatography.com/public/images/cheatography_logo.pdf}}
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    \vspace{-2pt}\large{\bf{\textcolor{DarkBackground}{\textrm{PTC - C4 (Explant Sterilization) Cheat Sheet}}}} \\
    \normalsize{by \textcolor{DarkBackground}{woozing} via \textcolor{DarkBackground}{\uline{cheatography.com/146689/cs/31799/}}}
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  \mymulticolumn{2}{p{5.377cm}}{\bf\textcolor{white}{Cheatographer}}  \\
  \vspace{-2pt}woozing \\
  \uline{cheatography.com/woozing} \\
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  \mymulticolumn{1}{p{5.377cm}}{\bf\textcolor{white}{Cheat Sheet}}  \\
   \vspace{-2pt}Not Yet Published.\\
   Updated 22nd April, 2022.\\
   Page {\thepage} of \pageref{LastPage}.
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  Measure your website readability!\\
  www.readability-score.com
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\begin{document}
\raggedright
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\begin{multicols*}{2}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Preparation of Stock Plants}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-Prior good care of stock plants may lessen the amount of contamination that is present on explants.} \tn 
% Row Count 2 (+ 2)
% Row 1
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-Transfer plants to a greenhouse to reduce endemic contaminants} \tn 
% Row Count 4 (+ 2)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-Plants grown in the field are typically more "dirty" than those grown in a greenhouse or growth chamber, particularly in humid areas like Malaysia.} \tn 
% Row Count 8 (+ 4)
% Row 3
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-Sources of contamination:} \tn 
% Row Count 9 (+ 1)
% Row 4
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{a) Overhead watering increases contamination of initial explants} \tn 
% Row Count 11 (+ 2)
% Row 5
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{b) Splashing soil on the plant during watering will increase initial contamination.} \tn 
% Row Count 13 (+ 2)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Correct Way to Water a Plant}}  \tn
\SetRowColor{LightBackground}
\mymulticolumn{1}{p{8.4cm}}{\vspace{1px}\centerline{\includegraphics[width=5.1cm]{/web/www.cheatography.com/public/uploads/woozing_1650620179_Screenshot 2022-04-22 173234.png}}} \tn 
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{p{0.8 cm} p{0.8 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Ways to have "Cleaner" Stock Plants}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{1. Force outgrowth of axillary buds} \tn 
% Row Count 1 (+ 1)
% Row 1
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-It is sometimes possible to harvest shoots and force buds from them in clean conditions.} \tn 
% Row Count 3 (+ 2)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-The forced shoots may then be free of contaminants when surface-sterilized in a normal manner.} \tn 
% Row Count 5 (+ 2)
% Row 3
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{2. Seeds may be sterilized and germinated in vitro to provide clean material.} \tn 
% Row Count 7 (+ 2)
% Row 4
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{3. Covering growing shoots for several days or weeks prior to harvesting tissue for culture may supply cleaner material.} \tn 
% Row Count 10 (+ 3)
% Row 5
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{4. Explants or material from which material will be cut can be washed in detergents and then placed under running water for 1 to 2 hours.} \tn 
% Row Count 13 (+ 3)
% Row 6
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{5. Remove surface contaminants by rinsing with sterilizing solutions} \tn 
% Row Count 15 (+ 2)
% Row 7
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{6. Treatment of stock plants with fungicides and/or bactericides is sometimes helpful.} \tn 
% Row Count 17 (+ 2)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Fungicides}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-pesticides that kill or prevent the growth of fungi and their spores} \tn 
% Row Count 2 (+ 2)
% Row 1
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-used to control fungi that damage plants, including rusts, mildews and blights} \tn 
% Row Count 4 (+ 2)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-used to control mold and mildew in other settings} \tn 
% Row Count 5 (+ 1)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
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\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Bactericide or Bacteriocide}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-substance which kills bacteria} \tn 
% Row Count 1 (+ 1)
% Row 1
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-it is disinfectants, antiseptics, or antibiotics} \tn 
% Row Count 2 (+ 1)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{p{0.8 cm} p{0.8 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Sterilization of Explants}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-Plants growing in the external environment are invariably contaminated with micro-organisms and pests} \tn 
% Row Count 3 (+ 3)
% Row 1
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-These contaminants are mainly confined to the outer surfaces of the plant, although, some microbes and viruses can even grow within the root.} \tn 
% Row Count 6 (+ 3)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-Most of the surface microbes do not form tight associations with the plant tissue.} \tn 
% Row Count 8 (+ 2)
% Row 3
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-They are easily removed from the explant by gentle rinsing, and the remaining are killed by surface sterilization.} \tn 
% Row Count 11 (+ 3)
% Row 4
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{} \tn 
% Row Count 11 (+ 0)
% Row 5
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-Place the explant in a 70\% ethyl alcohol (ethanol) solution prior to treatment with another disinfectant solution. } \tn 
% Row Count 14 (+ 3)
% Row 6
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-Conducting the sterilization process under vacuum results in the removal of air bubbles and provides a more efficient sterilization process.} \tn 
% Row Count 17 (+ 3)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{p{0.8 cm} p{0.8 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Sterilant}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{1. Oxidants} \tn 
% Row Count 1 (+ 1)
% Row 1
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{2. Active halogens} \tn 
% Row Count 2 (+ 1)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{3. Heavy metal poisoning} \tn 
% Row Count 3 (+ 1)
% Row 3
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{4. Powerful chemicals such as concentrated sulphuric acid may be used on seeds.} \tn 
% Row Count 5 (+ 2)
% Row 4
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{} \tn 
% Row Count 5 (+ 0)
% Row 5
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-cut surfaces should be protected} \tn 
% Row Count 6 (+ 1)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Common Explant Sterilization Agents}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{1. Sodium Hypochlorite} \tn 
% Row Count 1 (+ 1)
% Row 1
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{2. Ethanol or Isopropanol} \tn 
% Row Count 2 (+ 1)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{3. Calcium Hypochlorite} \tn 
% Row Count 3 (+ 1)
% Row 3
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{4. Mercuric Chloride} \tn 
% Row Count 4 (+ 1)
% Row 4
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{5. Hydrogen Peroxide} \tn 
% Row Count 5 (+ 1)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{p{0.8 cm} p{0.8 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{1. Sodium Hypochlorite}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-most frequent choice for surface sterilization} \tn 
% Row Count 1 (+ 1)
% Row 1
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-readily available and can be diluted to proper concentrations} \tn 
% Row Count 3 (+ 2)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-5.25\% sodium hypochlorite (laundry bleach) → 0.5 - 1.0\% sodium hypochlorite (sterilant)} \tn 
% Row Count 5 (+ 2)
% Row 3
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-Plant material is usually immersed in this solution for 10 - 20 minutes.} \tn 
% Row Count 7 (+ 2)
% Row 4
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-A balance between concentration and time must be determined empirically for each type of explant, because of phytotoxicity.} \tn 
% Row Count 10 (+ 3)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{p{0.8 cm} p{0.8 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{2. Ethanol or Isopropanol}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-powerful sterilizing agent but also extremely phytotoxic} \tn 
% Row Count 2 (+ 2)
% Row 1
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-plant material is typically exposed to it for only seconds or minutes} \tn 
% Row Count 4 (+ 2)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-more tender the tissue, the more it will be damaged by alcohol} \tn 
% Row Count 6 (+ 2)
% Row 3
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-Generally 70\% ethanol is used prior to treatment with other compounds.} \tn 
% Row Count 8 (+ 2)
% Row 4
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-Tissues such as dormant buds, seeds, or unopened flower buds can be treated for longer periods of time since the tissue that will be explanted or that will develop is actually within the structure that is being surface-sterilized.} \tn 
% Row Count 13 (+ 5)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{p{0.8 cm} p{0.8 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{3. Calcium Hypochlorite}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-obtained as a powder and must be dissolved in water} \tn 
% Row Count 2 (+ 2)
% Row 1
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-concentration that is generally used is 3.25 \%} \tn 
% Row Count 3 (+ 1)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-solution must be filtered prior to use since not all of the compound dissolve into solution} \tn 
% Row Count 5 (+ 2)
% Row 3
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-Calcium hypochlorite may be less injurious to plant tissues than sodium hypochlorite} \tn 
% Row Count 7 (+ 2)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{4. Mercuric Chloride}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-used only as a last resort} \tn 
% Row Count 1 (+ 1)
% Row 1
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-concentration of 0.1-1.0\% denatures proteins} \tn 
% Row Count 2 (+ 1)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-extremely toxic to both plants and humans and must be disposed of with care} \tn 
% Row Count 4 (+ 2)
% Row 3
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-Since mercury is so phytotoxic, it is critical that many rinses be used to remove all traces of the mineral from the plant material.} \tn 
% Row Count 7 (+ 3)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{5. Hydrogen Peroxide}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-concentration of 30\% is used, which is 10 times stronger than that obtained from pharmacy} \tn 
% Row Count 2 (+ 2)
% Row 1
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-useful for surface-sterilizing material while in the field} \tn 
% Row Count 4 (+ 2)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{p{0.8 cm} p{0.8 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Enhance Effectiveness of Sterilization}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-Surfactant/ wetting agent (e.g. Tween 20 or Triton-X) is frequently added to the sodium hypochlorite.} \tn 
% Row Count 3 (+ 3)
% Row 1
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-Wetting agents makes the surface of plant wet and repel the air, thus making the treatment more effective.} \tn 
% Row Count 6 (+ 3)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-The solutions that the explants are in are often shaken or continuously stirred.} \tn 
% Row Count 8 (+ 2)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Rinsing}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-After plant material is sterilized with one of the above compounds, it must be rinsed thoroughly with sterile water.} \tn 
% Row Count 3 (+ 3)
% Row 1
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-Typically three to four separate rinses are done} \tn 
% Row Count 4 (+ 1)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Use of Antibiotics and Fungicides in vitro}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-The use of antibiotics and fungicides in vitro is not very effective in eliminating microorganisms} \tn 
% Row Count 2 (+ 2)
% Row 1
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-These compounds are often quite phytotoxic.} \tn 
% Row Count 3 (+ 1)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{x{4 cm} x{4 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{If Sterilization is a Problem}}  \tn
% Row 0
\SetRowColor{LightBackground}
1. Seeds can be used instead & -Most seeds have hard outer coat that can withstand harsh and prolong sterilization procedure \tn 
% Row Count 5 (+ 5)
% Row 1
\SetRowColor{white}
 & -Germinate the seed in vitro to obtain axenic seedlings providing sterile meristems, leaves, roots and stem nodes for culturing \tn 
% Row Count 12 (+ 7)
% Row 2
\SetRowColor{LightBackground}
2. Grow plant under controlled environment & -Enclosed environment to prevent accumulation of dust \tn 
% Row Count 15 (+ 3)
% Row 3
\SetRowColor{white}
 & -Treatment with fungicide and to lesser extent antibiotics \tn 
% Row Count 18 (+ 3)
% Row 4
\SetRowColor{LightBackground}
 & -Clean organs can be collected later \tn 
% Row Count 20 (+ 2)
% Row 5
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{} \tn 
% Row Count 20 (+ 0)
% Row 6
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{*Axenic: state of a culture in which only a single species, variety, or strain of organism is present and entirely free of all other contaminating organisms} \tn 
% Row Count 24 (+ 4)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Explant Sterilization Process}}  \tn
\SetRowColor{LightBackground}
\mymulticolumn{1}{p{8.4cm}}{\vspace{1px}\centerline{\includegraphics[width=5.1cm]{/web/www.cheatography.com/public/uploads/woozing_1650644250_Screenshot 2022-04-23 001432.png}}} \tn 
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Phytotoxic}}  \tn
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-poisonous to plants% Row Count 1 (+ 1)
} \tn 
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}


% That's all folks
\end{multicols*}

\end{document}