Murashige & Skoog medium Murashige & Skoog (1962)

Inorganic Salt Formulation

Macronutrients (mg/l)


Ammonium nitrate (NH4NO3)
1,650mg/l
Calcium chloride (CaCl2*H2O)
440mg/l
Magnesium sulfate (MgSO4*7H2O)
370mg/l
Potassium phosphate (KH2PO4)
170mg/l
Potassium nitrate (KNO3)
1,900mg/l

Micronutrients (mg/l)


Boric Acid (H3BO3)
6.2mg/l
Colbalt chloride (CoCl2*6H2O)
0.025mg/l
Cupric Sulfate (CuSO4*5H2O)
0.025mg/l
Ferrous sulfate (FeSO4*7H2O)
27.8mg/l
Manganese sulfate (MnSO4*4H2O)
22.3mg/l
Potassiom Iodine (KI)
0.83mg/l
Sodium molybdate (Na2MoO4*2H2O)
0.25mg/l
Zinc Sulfate (ZnSO4*7H2O)c
8.6mg/l
Na2EDTA*2H2Oa
37.2mg/lb

Common Organic Additives


i-Inositol
100mg/l
Nicotinic Acid
0.5mg/l
Pyridoxine*HCl
0.5mg/l
Thiamine *HCl
0.1mg/l
IAA
1-30mg/l
Kinetin
0.04-10mg/l
Glycine (recrytallized)
2.0g/l
Edamine
1.0g/l
Sucrose
20g/l
Agar
10g/l

a= Originally printed as Na2EDTA
b= Originally printed as 37.3 mg/l anhydrous form
c= Originally printed as ZnSO4*4H2O

Hodgkin and Lyon's Medium ( Hodgkin and Lyon 1986 ) ; standardized for brassica oleracea :

Hodgkin and Lyon's Medium ( Hodgkin and Lyon 1986 ) ; standardized for brassica oleracea :

Socrose 580 mM ( ca. 20% )

Boric acid 1.62 mM ( ca. 100mg/L )

Calcium nitrate 1.69 mM ( ca. 400mg/L )

Magnesium sulfate 0.84 mM ( ca. 200mg/L )

Potassium nitrate 0.99 mM ( ca. 100mg/L )

TAPS 20 mM ( ca. 4.86g/L )

( pH of medium adjusted to 8 with 0.1 N NaOH )

Roberts' Medium ( Roberts et al. 1983 ) ; standardized for brassica oleracea :

Roberts' Medium ( Roberts et al. 1983 ) ; standardized for brassica oleracea :

Socrose 20%

Boric acid 10mg/L

Calcium chloride 362mg/L

Potassium nitrate 100mg/L

Tris 60-130mg/L

brewbaker and kwack's medium

brewbaker and kwack's medium (brewbaker and kwack 1963) ; found suitable for some 86 species :

Socrose 10%

Boric acid 100mg/L

Calcium nitrate 300mg/L

Magnesium sulfate 200mg/L

Potassium nitrate 100mg/L

百合花粉發芽試驗

為什麼需要做花粉活性試驗??

育種時，親本的花期無法配上時，就需將父本的花粉收集保存，等優良母本花出現再行授粉，但要如何知道儲存的花粉是否還有活力，授粉是否還有效，就需要做花粉活力試驗。

要知道花粉活性大略有下例方法

1. 染色法:
利用染劑將花粉內酵素、DNA、RNA染色，來做判斷，此法最快最省時
2. 體內萌發
柱頭授粉壓片於螢光顯微鏡觀察，針對不易體外萌發的花粉，這方法最準確，可看出花粉對柱頭親和性，胚珠是否授精
3. 花粉發芽試驗
將花粉置於養液下培養，此方試較省成本，最好的好處看到的花粉是活的狀態，你可看到花粉正在成長

有機會再和各位介紹上面二種方法，今天先介紹百合花粉發芽試驗

配製經過修改的BK 培養基(modified BK medium)，混合花粉後，就可開始培養了



部署完成



較黑的花粉未發芽，白色的花粉已發芽


92% 的發芽率

形成隔膜



細胞核(nuclei) 這應該是精核(sperm cell)

百合花粉原生質流

http://picasaweb.google.com.tw/changzhanrong/SIHuXG#5290412153027580594

這次實驗鐵砲百合做到90% 以上發芽率，準確度很高，可檢測花粉有無問題，測試儲存的條件是否夠好，做為下次儲藏參考，一搬來說花粉還有二成有活力就夠了，但活力愈高，當然是愈好。