Table 1.
Disinfection procedures of tissues for in vitro use of Vitex species (alphabetical listing).
| Species | Disinfection protocol | References |
|---|---|---|
| V. agnus-castus | After removal of testa by mechanical treatment → 1.25% NaOCl (5% available chlorine) 20 min → SDW 4–5× | [22] |
| V. agnus-castus | Shoot tip/nodal explant → RTW → 0.05% Bavistin® 5 min → SDW 3–4 × → SDW + 2–3 drops Tween-20 10 min → SDW 3–4 × → 70% EtOH 30 s → SDW 3–4× | [16] |
| V. agnus-castus | Apical bud or stem with nodes from seedlings → 75% EtOH 15 s → 0.1% HgCl2 3–5 min → SDW 5–6× | [59] |
| V. doniana | Second pair of young leaves from 2–4 year old tree → cleaned with cotton wool soaked in liquid soap → 0.5% fungicide (Ridomil®) + 2 drops of Tween 20 1 h → 70% EtOH 30 min → SDW 2 × → CaCl2 (1, 1.5, 2%) + 2–3 drops of Tween 20 (duration NR) → 2% CaCl2 30 min → SDW 2 × → quick dip in 70% EtOH → 2% CaCl2 15 min → SDW 2 × → 70% contamination-free cultures obtained | [29] |
| V. leucoxylon | Explants → 10% NaOCl 10 min → SDW 4–5× | [25] |
| V. negundo | Young shoots → 1% Teepol® (detergent) → 0.1% HgCl2 (2 min) → repeated wash with SDW (node with one axillary bud 1 cm) | [108] |
| V. negundo | Shoot apices and nodal segments → RTW 45 min → 5% Laboline® (detergent) + 7% NaOCl (7–10 min) → 5–7 × SDW → 0.1% HgCl2 8 min → SDDW 6–7× | [80] |
| V. negundo | Shoot apices and nodal segments → RTW → 1% Teepol® 10 min → RTW → 80% EtOH 30 s → 0.1% HgCl2 3 min→ SDW 3× | [102] |
| V. negundo | Internode segment (1.0–1.50 cm) from 10-year-old plant → RTW → 1%Teepol® 10 min → RTW → 80% EtOH 30 s → 0.1% HgCl2 3 min | [103] |
| V. negundo | Shoot apices and nodal segments → RTW → 1% Teepol® 10 min → RTW → 50% EtOH 30 s → 0.1% HgCl2 3 min→ SDW 3× | [104] |
| V. negundo | Actively growing healthy shoot with 3–4 nodes from adult plant (0.5–1.0 cm node with dormant axillary bud) → RTW 30 min → 5% Laboline® 7–8 min → DW → 0.05% HgCl2 20 min → SDW 5–6× | [24] |
| V. negundo | Young leaves (< 3 months) → 2% detergent + 2.5% commercial bleach + 0.01% NaOCl → 70% EtOH → SDW | [67] |
| V. negundo | Shoot tip/nodal explant with one dormant axillary bud (1.0–1.50 cm) from 6-month-old plant in March → RTW 30 min → 10% Laboline 10 min → washed thoroughly with SDW → 0.1% HgCl2 5 min → SDDW 6–7× | [77] |
| V. negundo | Explants → RTW → 5% Extran® (detergent) 10 min → 1% Bavistin (fungicide) 20 min → SDDW → 0.05% HgCl2 4 min → several rinses in SDDW | [107] |
| V. negundo | Young shoots → RTW 30 min → 5% Laboline® (2007, 2008, 2013b) or Teepol® (2011) 10 min → SDW 3–4 × → 0.1% HgCl2 4 min (2007, 2013) or 5–7 min (2008, 2011) → SDW 5–8× | [3], [5], [7], [6] |
| V. negundo | Shoot tips (1.0–1.5 cm) → 0.5% Tween-20 → 70% EtOH 10 s → 0.1% HgCl2 3 min → SDW 4–5× | [106] |
| V. negundo | Young leaves from 3–5-month-old plant → RTW → 5% Tween-20 10 min → 70% EtOH 10–15 s → SDW 5–10 min → 0.1% HgCl2 2–3 min → SDW 4–5× | [49] |
| V. negundo | Young shoots → RTW → 0.1% HgCl2 1 min → SDW 3–4× | [50] |
| V. negundo | Explants → 5% Teepol® 10–15 min → 0.1% HgCl2 5 min → SDW 3× | [68] |
| V. negundo | Twigs → RTW → DW → shoot tips excised → 0.1% HgCl2 7 min → SDW 3–5× | [47] |
| V. negundo | Explants → RTW → detergent 30 min → 0.1% HgCl2 7 min → SDW 5× | [48] |
| V. negundo | Explants → water 10–15 min → 0.1% HgCl2 7–10 min → SDW 5–7× | [85] |
| V. negundo | Young stems 15–20 mm → RTW 30 min → Tween-80 → SDW 4× | [26] |
| V. negundo | Twigs → RTW → 1% Savlon® + liquid soap 5–10 min with shaking → SDW 3–4 × → 70% EtOH < 1 min → 0.1% HgCl2 5–7 min → SDW 4–5× | [75] |
| V. negundo | 0.1% Bavistin® 10–15 min → 0.1% antibiotics (streptomycin and tetracycline) 5–10 min → 0.1% HgCl2 3–4 min → SW 5–6 × → 0.1% AA + 0.05% CA | [78] |
| V. negundo | RTW → Tween-20 (2 drops/100 ml) → 0.1% HgCl2 3–5 min → dip in 70% EtOH → SDW 4–5× | [41] |
| V. negundo | RTW 3 min → cut into 3–5 cm sections with 2–3 internodes → 2 g/l Blitox® (fungicide) + Tween-20 45 min → DW 3 × → SDW 1 min → AA + CA 2 min → SDW 3 × → 70% EtOH 30 s → SDW 2× → 0.12% HgCl2 + 1 drop Tween-20 8 min → SDW 3× | [79] |
| V. negundo Dhaka 1205 | Shoot tip/nodal explant → RTW → detergent 30 min → 70% EtOH 1 min → 0.1% HgCl2 5 min → SDW 5× | [2] |
| V. rotundifolia | Nodal explant → RTW → 70% EtOH 10 s → NaOCl (2% active chlorine) 15 min → SDW 3× | [72] |
| V. trifolia | 1-cm long nodal explants → RTW → 0.1% Laboline® 10 min → 70% alcohol 5 min → 0.1% HgCl2 5 min → several rinses in DW | [44] |
| V. trifolia | RTW → Bavistin® + Tween-20 10–15 min → 70% EtOH 1 min → 0.1% HgCl2 4 min → SDDW | [15] |
| V. trifolia | 5–7 cm long young shoots → RTW 20 min → 5% Laboline® 5 min → DW 3–4 × → 0.1% HgCl2 4 min → SDW repeatedly | [10], [11], [12], [13] |
| V. trifolia | Shoot tips → RTW 10 min → 5% Tween-20 15 min → DW → 0.5% NaOCl 5 min → DDW → 0.01% HgCl2 10 min → 0.1% HgCl2 5 min → Bavistin® 30 min → DDW 1× | [64] |
| V. trifolia | 5–7 cm long shoots → 0.1% HgCl2 20 min → rinse 4–5 × in SDW | [81] |
AA, ascorbic acid; CA, citric acid; DDW, double distilled water; DW, distilled water; EtOH, ethyl alcohol (ethanol); HgCl2, mercuric chloride; NaOCl, sodium hypochlorite; NR, not reported; RTW, running tap water; s, second(s); SDW, sterilized (by autoclaving) distilled water; SDDW, sterilized (by autoclaving) double distilled water; SW, sterile water.