1. 百合2n花粉的發生 Occurrence of 2n-Pollen in Lilium
學生：莊 畫 婷
Hua-Ting, Chuang
學號：
日期：2008/03/19

2. Outline Introduction Detection of 2n-pollen in Lilium
Conclusion
Detection of 2n-pollen in Lilium
Mechanisms of 2n-pollen in Lilium
Utilization of 2n-pollen in Lilium

3. Outline Introduction Detection of 2n-pollen in Lilium
Mechanisms of 2n-pollen in Lilium
Utilization of 2n-pollen in Lilium
Conclusion

4. 2n-pollen (or unreduced pollen ) originate due to deviating meiosis in plants.
The process that leads to 2n-pollen formation is called meiotic nuclear restitution.

5. Year
Investigator
Event
1927
Rosenberg
Meiotic nuclear restitution was
first proposed.
1982
Asano
In lily, unreduced gametes (2n-
pollen) are found in some wide
interspecific hybrids.

6. n-pollen 2n-pollen Metaphase Product Normal meiosis Plant 2n=2x=4
Meiotic nuclear restitution
2n-pollen
(Ramanna and Jacobsen, 2003; Veilleux, 1985)

7. Outline Introduction Detection of 2n-pollen in Lilium
Mechanisms of 2n-pollen in Lilium
Utilization of 2n-pollen in Lilium
Conclusion

8. Detection of 2n-pollen in Lilium
Progeny
Morphological
Flow cytometry
Microsporogenesis
Meiotic observation

9. The unexpected occurrence of polyploid progeny from 2x-2x, 4x-2x or 2x-4x hybridization has usually been the first indication of functional 2n-gametes.
(Ramanna and Jacobsen, 2003; Veilleux, 1985)

10. Percentage of aneuploids
Progeny
Table 1. Chromosome number in progeny obtained by cro sses between Lilium ‘Grand paradiso’ and 2n-pollen of L. ‘Mona’.
Cross combinations
Chromosome number
Percentage of aneuploids
Seed parents
Pollen parents 24 40 41 42 43 44 45 46 47 48
Gran paradiso
Mona 1 5 10 6 7 3 9
80.8
4x × 2x 4x
利用雜交後代發生
(Akutsu et al., 2007)

11. Morphological
補非百合的資訊，如2n-pollen為n-pollen的幾倍大。
Fig. 1. Comparison of n and 2n-pollen from Lilium ‘Alaska’ (2n = 24) (a) pollen. (b) 2n pollen. Bar=100µm .
(Akutsu et al., 2007)

12. Pollen grain diameter (µm)
Morphological
Pollen grain diameter (µm)
No. of pollen grains
(a)
(b)
3.5%
2n-pollen
28.1%
2n-pollen
Fig. 2. Distribution of pollen grain size from Lilium inter specific hybrids . (2n = 24).
(Van Tuyl et al., 1989)

13. Flow cytometry
(a)
(c)
(b)
Number of nuclei
Relative DNA content
Fig. 3. Flow cytometric analyses of root tip and pollen from Lilium interspecific hybrids (a) Root (from lily 2n=2x=24) (b) Pollen (from lily 2n=2x=24) (c) Root (from lily 2n=4x=48).
(Van Tuyl et al., 1989)

14. Microsporogenesis
(a)
(b)
Unbalanced chromosome distribution. This type of disjunction occurs the most commonly druing microsporogenesis in distantly related inter-specific hybrids. Unbalanced chromosome distribution. This type of disjunction occurs the most commonly druing microsporogenesis in distantly related inter-specific hybrids.
Fig. 4. Microsporogenesis from Lilium interspecific hybrid (a) Triad shows a clear division of three nuclear formations by succe ssive cytokinesis (b) Sporads after tetrads stage, show dyads (D), triads (T) and tetrads.
(Lim et al., 2004)

15. Table +1. Frequency of types sporocytes at tetrad stage from Lilium hybrids.
Monad
Dyad
Triad
Tetrad
Others
Total
Revival －
1(1)*
3(2)
124(97)
128(100)
8(5)
113(64)
18(10)
29(16)
176(100)
11(6)
97(53)
23(13)
45(25)
5(3)
181(100)
Normal
2n-Pollen
* The percentage is given in brackets.
(Lim et al., 2004)

16. Meiotic observation
(a)
(b)
(c)
Fig. 5. Meiosis and meiotic nuclear restitution from Lilium interspecific hybrid and (a) Balanced movement of homoeologous chromosomes at late ana phase I. ( ) (b) Unbalanced chromosome distribution. ( ) (c) Restitution nucleus formation in pollen mother cells without cytoki nesis (arrows; ).
Unbalanced chromosome distribution. This type of disjunction occurs the most commonly druing microsporogenesis in distantly related inter-specific hybrids. Unbalanced chromosome distribution. This type of disjunction occurs the most commonly druing microsporogenesis in distantly related inter-specific hybrids.
(Barba-Gonzalez et al., 2005 Lim et al., 2004)

17. Meiotic observation 24 I 4II+16I
(1a) Metaphase I showing 3 bivalents (arrowheads) and 18 univalents.
染色體配對異常發生
(a)
(b)
Fig. 6. Chromosome pairing at Metaphase I stage from Lilium interspecific hybrid
(Barba-Gonzalez et al., 2004)

18. Detection of 2-n pollen in Lilium
1.The unexpected occurrence of polyploid progeny from hybridization.
2.The larger (>90μm) grains represent 2n pollen ( %).
3.2n-Pollen from diploid genotypes gave rise to 1C and 2C peak by flow cytometry, can be related to haploid and diploid pollen.

19. Detection
4.High frequency of dyads (53-64%) was formed after microsporogenesis.
5.Disturbed chromosome pairing.
6.Restitution nucleus formation in pollen mother cells without cytokinesis or unbalanced chromosome distribution.

20. Outline Introduction Detection of 2n-pollen in Lilium
Mechanisms of 2n-pollen in Lilium
Utilization of 2n-pollen in Lilium
Conclusion

21. Prophase I
Metaphase I
Anaphase I
Cytokinesis I
Metaphase II
Anaphase II
Cytokinesis II
Product
Normal
FDR
IMR
SDR
PMR
理論上，FDR在減數分裂第一期是不具有同源染色體配對的情形。
(Ramanna, 1979)FDR include mainly non-sister chromatids of each homologous pair of chromosomes.
SDR the sister chromatid are included in the same gametes.
Mother cell 2n=2x=4
Fig. 7. Schematic representation of the meiotic process and restitution mechanisms in microsporogenesis.
(Lim et al., 2001; Lim et al., 2004; Karlov et al., 1998; Ramanna and Jacobsen, 2003; Veilleux, 1985)

22. All univalent or a part univalent
All bivalent
Fig. 8. Schematic representation of two possible types of meiotic nuclear restitution in a diploid interspecific hybrid (2n=2x=4) (a) FDR: First division restitution (b) SDR: Second division restitution.
(Lim et al., 2001)

23. A part of bivalents
Bivalents
Univalents
Fig. 9. Genomic in situ hybridization to the chromosomes at Metaphase I stage from Lilium interspecific hybrids (L. longiflorum × Asiatic, LA) for FDR Red fluorescence: Asiatic Yellow fluorescence: L. lolngiflorum Bar= 10 µm.
(Lim et al., 2001)

24. All bivalents
Fig. 10. Chromosome pairing at Metaphase I stage from Lilium interspecific hybrid (2n=24) for SDR.
(Lim et al., 2004)

25. Configurations / cells
Table 2. Frequency of chromosome association at metaphase I stage in night hybrids of Lilium.
Hybrids
No. of cells observed
Configurations / cells
Mean frequency
12II
11II
10II
9II
8II
7II
6II
5II
4II
3II
2II
1II
0II 0I 2I 4I 6I 8I
10I
12I
14I
16I
18I
20I
22I
24I
Revial
125
115 8 2 －
11.8II+0.2I 99 95 3 1
11.9II+0.1I
118
108 6 4 32 5 12
3.7II+16.6I 87 13 20 28 10
2.1II+19.8I
882111
117 19 34 39 23
1.5II+21.0I
148 17 35 26 31
2.6II+18.8I
191 7 29 64 61 30
1.6II+20.8I
296 63 91
1.25II+21.49I 50 14
1.06II+21.88I
Normal, II→92%
SDR, II→92-96 %
FDR，II→0%
由會產生2N-花粉的百合，來觀查花粉形成的機制，79的兩個百合應該是屬於SDR型，其他的應該是屬於FDR或IMR型，MI期，染色家構形異常，理論上，完全沒有配對時，可推測是FDR型發生機制的2N-GAMETES，
但實際上，很少有這種情形，因此很難辨別是FDR或IMR的機制
(Van Tuyl et al., 2005)

26. All univalent or a part univalent
A part bivalent or a part univalent
觀察中期以外的染色體動態，相當困難。
Fig. 11. Schematic representation of three possible types of meiotic nuclear restitution in a diploid inter specific hybrid (2n=2x=4) (a) FDR: First division restitution (b) SDR: Second division restitution (c) IMR: Indeterminated meiotic restitution.
(Lim et al., 2001)

27. Configurations / cells
Table 2. Frequency of chromosome association at metaphase I in eight hybrids of Lilium.
Hybrids
No. of cells observed
Configurations / cells
Mean frequency
12II
11II
10II
9II
8II
7II
6II
5II
4II
3II
2II
1II
0II 0I 2I 4I 6I 8I
10I
12I
14I
16I
18I
20I
22I
24I
Revial
125
115 8 2 －
11.8II+0.2I 99 95 3 1
11.9II+0.1I
118
108 6 4 32 5 12
3.7II+16.6I 87 13 20 28 10
2.1II+19.8I
882111
117 19 34 39 23
1.5II+21.0I
148 17 35 26 31
2.6II+18.8I
191 7 29 64 61 30
1.6II+20.8I
296 63 91
1.25II+21.49I 50 14
1.06II+21.88I
FDR or IMR？
由會產生2N-花粉的百合，來觀查花粉形成的機制，79的兩個百合應該是屬於SDR型，其他的應該是屬於FDR或IMR型，MI期，染色家構形異常，理論上，完全沒有配對時，可推測是FDR型發生機制的2N-GAMETES，
但實際上，很少有這種情形，因此很難辨別是FDR或IMR的機制
(Van Tuyl et al., 2005)

28. Table 3. Origin and parentage of Lilium interspecific hybrids and BC1.
2n-pollen
Genome typea
Accession number
Female parent × male parent
F1 hybrids
LA1~3
,52,69
L. longiflorum Gelria × Asiatic hybrid Whilito
BC1 progeny
ALA1
Asiatic hybrid ×
ALA2~4
,2,3
Asiatic hybrid Montreux ×
ALA5~6 ,2
Asiatic hybrid Puccini ×
ALA7~8 ,2
Asiatic hybrid Meribel ×
a L and A, L. longiflorum and Asiatic hybrid, respectively.
(Lim et al., 2001)

29. Fig. 12. Triploid (2n=3x=36) chromosome constitution of BC (ALA1) lily, derived through the functioning of a 2n pollen that originated through IMR.
其中有27條A基因組，9條L基因組，兩者皆屬於奇數型式。箭>表示有5對染色體發生重組，箭頭→表示中心粒的位置。
白字代表「」，黃字代表「」。
(Lim et al., 2001)

30. Table 4. Number of chromosomes and chromosome consti tution from Lilium interspecific hybrids and BC1.
Genome
typea
Somatic chromosome number (2n)
Chromosome
constitutiona
Number of cross-over break-points
Number of recombinant chromosomes
Nechanism of the 2n-gamete involved L A
L/Ab
A/Lb
Total F1
LA1 24 12 －
LA2
LA3
BC1
ALA1 36 9 27 10 1 4 5
IMR
ALA2 7 2 3
FDR
ALA3
ALA4 48
ALA5
ALA6
ALA7
ALA8
odd
由回交後代鑑定，親本染色體分配呈現奇數，即為IMR的型式。
a L and A, L. longiflorum and Asiatic hybrid, respectively.
b L/A and A/L, Recombinant chromosomes of L. longiflorum chromosome (centromere) with Asiatic chromosome
and Asiatic hybrid chromosomeS (centromere) with L. longiflorum chromosome, respectively.
(Lim et al., 2001)

31. The chromosome of parent is even number.
Porphase I
Metaphase I
Anaphase I
Cytokinesis I
Metaphase II
Anaphase II
Cytokinesis II
Product
Normal
FDR-
lily type
The chromosome of parent is even number.
FDR
IMR
理論上，FDR在減數分裂第一期是不具有同源染色體配對的情形。
(Ramanna, 1979)FDR include mainly non-sister chromatids of each homologous pair of chromosomes.
SDR the sister chromatid are included in the same gametes.
Mother cell 2n=2x=4
The chromosome no. of parent is odd.
Fig. +1. Schematic representation of the meiotic process and restitution mechanisms in microsporogenesis.
(Lim et al., 2001; Lim et al., 2004; Karlov et al., 1998; Ramanna and Jacobsen, 2003; Veilleux, 1985)

32. FDR
Failure of chromosome pairing (form all univalent or a part univalent) at meiotic I stage.
Without the second division, abnormal cytokinesis and give rise to dyads instead of tetrads.
特性：1、減數分裂初期有些同源染色體有配對，有些沒有配對情形；2、姊妹染色分體平均分離；3、配子平均分離，呈現異質結合性，姊妹染色分體不存在於同一細胞中。
Equational division of sister chromatids.
The chromosome of parent is even number (to compare IMR).

33. SDR Normal chromosome pairing (All bivalent) at meiotic I stage.
Abnormal cytokinesis at meiotic II stage.
Sister chromatids move to the same daughter cell.
特性：1、減數分裂第一中期有些同源染色體有配對情形；2、有同源染色體分離情形；3、姊妹染色分體存在於同一細胞中。

34. IMR Display a mixture of FDR and SDR.
Failure of chromosome pairing (A part univalent) at meiotic I stage.
A part sister chromosome move to the same daughter cell.
The parental genomes are present in odd numbers.
特性：1、減數分裂初期有些同源染色體有配對，有些沒有配對情形；2、姊妹染色分體平均分離；3、配子平均分離，呈現異質結合性。

35. Outline Introduction Detection of 2n-pollen in Lilium
Mechanisms of 2n-pollen in Lilium
Utilization of 2n-pollen in Lilium
Conclusion

36. Utilization
2n-Pollen have been demonstrated to function in fertilization in many plants.
2 n－花粉已經被利用在許多植物中的繁殖。
(Veilleux, 1985)

37. Fig. 13. 2n-Pollen germination from Lilium interspecific hybrid 79418-2.
(Lim et al., 2004)

38. Table 5. Pollen germination of n and 2n-gametes from Lilium ‘Revial’ and interspecific hybrid
Genotype
Pollen type
No. of pollen
Germinated pollen
Percentage
In group
In total
Revial n
251
173
68.9
68.1 2n 3
All
254 －
130 41
31.5
19.1 85 54
63.5
25.1
215 95
44.2
Normal
2n-Pollen
2n-pollen具有發芽能力
(Lim et al., 2004)

39. Chromosome recombinant
在具有2n-pollen的OA雜交百合，觀察其減數分裂中期之情形，有重組的產生，增加遺傳多樣性，有利育種。[並在BC1的分析上得知，2N-POLLEN是具有實用性的]。
Fig. 14. Genomic in situ hybridization to the Chromosome at Post-Metaphase I stage from Lilium interspecific hybrid (with 2n-pollen) Post-Metaphase I stage in which sister chromatids of each chromo some are clearly visible, as are the recombinant chromatids in three pairs (arrow heads) and the double strand crossover event (arrows) Bar: 10μm.
(Barba-Gonzalez et al., 2005)

40. Chromosome recombinant
其中有27條A基因組，9條L基因組，兩者皆屬於奇數型式。箭>表示有5對染色體發生重組，箭頭→表示中心粒的位置。
白字代表「」，黃字代表「」。
Fig. 15. Genomic in situ hybridization to the Chromosome from BC1 (ALA1) of Lilium (2n=3x=36).
(Lim et al., 2001)

41. Number of recombinant chromosomes
Table 6. Genome composition and number of recombinant chromo somes in progenies obtained from the cross of N2O trea ted OA lily hybrid to Asiatic (A) parent.
Genotype
Cross
Ploidy
Genome composition
Number of recombinant chromosomes
O (O/A)
A (A/O)
AA×OA 3x 10
26(2) 2 4x
12(1)
36(1) 12 24 36 13 35
OA×AA 1
重組的情形
(Barba-Gonzalez et al., 2006b)

42. Table +2. Number of chromosomes and chromosome consti tution from Lilium interspecific hybrids and BC1.
Genome
typea
Somatic chromosome number (2n)
Chromosome
constitutiona
Number of cross-over break-points
Number of recombinant chromosomes
Nechanism of the 2n-gamete involved L A
L/Ab
A/Lb
Total F1
LA1 24 12 －
LA2
LA3
BC1
ALA1 36 9 27 10 1 4 5
IMR
ALA2 7 2 3
FDR
ALA3
ALA4 48
ALA5
ALA6
ALA7
ALA8
由回交後代鑑定，親本染色體分配呈現奇數，即為IMR的型式。
a L and A, L. longiflorum and Asiatic hybrid, respectively.
b L/A and A/L, Recombinant chromosomes of L. longiflorum chromosome (centromere) with Asiatic chromosome
and Asiatic hybrid chromosomeS (centromere) with L. longiflorum chromosome, respectively.
(Lim et al., 2001)

43. Despite these advantages, however, there is a limitation for routinely using 2n-pollen in Lilium breeding.
Because the genotypes that produce 2n-pollen spontaneously are selected through laborious process.
1.儘管2n-花粉的百合具有發芽組力及基因重組的遺傳多樣性之優點，然而，有限制為通常在農作物飼養中使用 2 n 配偶子 。
2.因為在育種過程中，選拔自然生產 2 n 花粉的基因型的個體來做為親本，相當費時費力。
3. 發展誘導 2 n花粉的方法將會是必要的。
In order to overcome this difficulty, it would be essential to develop methods of inducing 2n-pollen.
(Barba-Gonzalez et al., 2006)

44. Table 7. The effects of heat treatment on Lilium OA hybrids.
Crossing number
Parentage
Pollen germination* (%) (range%)
Oriental Hybrid
Asiatic Hybrid
Heated**
Unheated
‘Romero Star’
‘Connecticut King’ －
31.4 (0-75)
‘Bel Paso’
‘Gran Sasso’
2.0 (n.a.)
‘Pesaro’
4.9 (0-10)
16.6 (0-100)
‘Acapulco’
‘Sancerre’
35.1 (0-90)
23.3 (0-60)
‘Expression’
‘Au Revoir’
25.0 (n.a.)
‘Mero Star’
2.6 (0-20)
7.6 (0-75)
0.53 (0-20)
‘San Marco’
37.5 (20-50)
6.0 (0-40)
‘Bernini’
1.9 (0-25)
‘Tenerife’
‘Lanzorote’
2.1 (0-30)
‘Sissi’
‘Mirella’
12.2 (0-60)
24.4 (0-75)
Up 66%
Up 7.0%
*Plants of four normally sterile F1 hybrid OA genotypes were grown in a phytotron for 6 weeks and
熱處理後，計算2n-pollen發芽率， 和 經熱處理後，2n-pollen發芽率上升。
n.a. = not available; －= genotype was not present. *The pollen germination percentage was scored counting only the large germinated 2n pollen grains. **10℃/8h→30℃/8h under artificial lighting and 10℃/4h→30℃/4h in the dark, creating day/night ratio of 16:8 h.
(Lokker et al., 2005)

45. Table 8. The effects of colchicine treatment on Lilium oriental.
Varieties
Concentration (%)
Flower buds of treament
Percentage of death (%)
Percentage of mutation (%)
‘Con.Amore’
0.00
300
0.0
0.02
12.7
0.05
37.3
4.7
0.10
49.3
9.5
0.20
81.3
1.2
‘Acapulco’
5.7
2.3
16.3
5.0
33.3
25.8
47.0
11.3
*Lilium were treated by different concentration colchicine 2hen the young flower buds were about 0.5cm length for the production of 2n-gametes.
The treating young flower buds were wrapped with a cotton wool soaked in colchicine and last for 3 days /14-18℃
利用秋水仙素來阻斷紡錘絲的形成，進而產生2n-pollen，2n-pollen的萌芽情形。
*The treating young flower buds (0.5cm length) were wrapped with a cotton wool soaked in
colchicine and last for 3 days.
(Wu et al., 2007)

46. Pollen grain diameter (µm)
No. of pollen grains
2n-pollen
利用N2O可以促進2N-POLLEN的形成。
(N2O:O2→5:1)pressure was at 6 atm.
Fig. 16. Distribution of pollen grain size from Lilium Lilies were treated with N2O gas for 24 h (a) ‘Alaska’ (2n = 24) (b) ‘Roma’ (2n = 24).
(Akutsu et al., 2007)

47. Fig. 17. Germination of 2n pollen from Lilium OA hybrid 951502-1
Fig. 17. Germination of 2n pollen from Lilium OA hybrid (a) Untreated (0 h treatment) (b) After 48 h of N2O treat ment. Bar: 0.5 cm.
2N花粉發芽情形。
(Barba-Gonzalez et al., 2006b)

48. No. of ovaries developed Ploidy level of seeds (%)
Table 9. Interploidy crosses between diploid and tetraploid lily varieties and crosses between tetraploids and N2O treated pollen.
Female
Male
No. of pollinated
No. of ovaries developed
No. of seeds obtained
Ploidy level of seeds (%)
patents
Diploid
Tetraploida
Diploid ×
diploid
Roma
Mona 5
305 (66)
100 (n=42)
Tetraploid×
tetraploid
Gran Paradiso
Avignon
240 (48)
100 (n=18)
Toreso
135 (27)
100 (n=30) －
N2O-treated pollen 11 8
175 (22)
100 (n=69) 4 3
19 (6)
100 (n=19)
17 (3)
100 (n=12)
本表說明了使用誘導劑可以增加雜交成功率，但卻顯示沒用者，則無雜交成功的情形。
a Ploidy of mature seeds was examined by flow cytometry; n the number of seeds examined.
(Akutsu et al., 2007)

49. Table 10. Number of embryos obtained after N2O treatment of sterile F1 Oriental × Asiatic (OA) lily hybrids and triploid BC (AOA) interspecific lily hybrids.
Genotype
Cross
#pollinations
#embryos
♀ × ♂
042923
AA* OA 19
179
042924 AA 3 35 4 1 6 28
052121
980072
AOA 14
052124
031028 20 10 32
950181 LD 2
051064
012046
利用N2O處理F1♂，來增加2N-pollen的形成，再進行回交，皆可克服不稔性，並獲得後代BC1。(少了一張沒處理誘導劑的結果或直接使用2n-pollen的結果)
*AA = Asiatic, OA = F1 hybrid Oriental × Asiatic, LD = L. longiflorum × L. dauricum, AOA = Triploid hybrid obtained by the cross (Oriental × Asiatic) × Asiatic.
(Barba-Gonzalez et al., 2006a)

50. Pollen germination (Range %)
Table 11. Time effect on the pollen germination of diploid Asiatic (AA) and Oriental (OO) cultivars and diploid OA hybrids of Lilium 2n pollen producers.
Code (Genome)
Flower no.
Pollen germination (Range %)
Fresh
1-day old
Pollyanna (AA) 12
95-100
Sorbonne (OO) 15
(OA) 17
85-100
0-<5
(OA) 20
0-40
2n花粉雖具發芽力，但開花後一天就失效，是在應用上需注意的。
☆The application of 2n-pollen needs to notice the fresh.
(Barba-Gonzalez et al., 2005)

51. Conclusion Detection of 2n-pollen in Lilium:
1. Unexpected occurrence of polyploid progeny.
(ex. 4x-2x→4x)
2. 2n-Pollen grains is larger than n pollen.
3. 2n-Pollen from diploid genotypes gave rise
to 2C peak (relative DNA content of 2n-pollen
more than n pollen) by flow cytometry.
4. High frequency of dyads.
5. Disturbed chromosome pairing.
6. Abnormal cytokinesis.

52. Conclusion Mechanisms: FDR IMR SDR Failure Chromosome pairing
(All bivalent)
Failure
Normal
Second division X V
(but abnormal cytokinesis)
Sister chroma-
tids to daugher
cell
Equational into two daughter cell.
A part move to same daughter cell.
Move to same daughter cell.
Chromosome of
parent
Even number
Odd number

53. Conclusion Utilization of 2n-pollen in Lilium:
Lilies sterile F1 hybrids can be readily used by 2n-pollen.
Utilization of 2n-pollen in Lilium:
Interspecific recombination can be accomplished for generating genetic variation in the F2 and BC1 progenies.
1、 透過2N花粉的利用，使具有不稔性的百合，再度成為有利用價值的親本。
2、2N花粉配子具有種間雜交染色體互換重組的能力，可以產生更多的遺傳變異，增加遺傳岐異度，對育種是有利的。
3、但並目前仍儘有少數的雜交種可以產生較多的2n-花粉，為使育種更順利，可利用熱休克、秋水仙素或氮氣處理，以增加2n花粉的產生。
4、另外在利用上要注意花粉的新鮮度 (開花一天後的花粉發芽率<5%)，才能使2n花粉順利發芽。

54. Conclusion Utilization of 2n-pollen in Lilium:
Inducement of 2n-pollen in lily by heat, colchicine or N2O treatment.
Utilization of 2n-pollen in Lilium:
Induced higher production of 2n-pollen will certainly increase the breeding parents.
1、 透過2N花粉的利用，使具有不稔性的百合，再度成為有利用價值的親本。
2、2N花粉配子具有種間雜交染色體互換重組的能力，可以產生更多的遺傳變異，增加遺傳岐異度，對育種是有利的。
3、但並目前仍儘有少數的雜交種可以產生較多的2n-花粉，為使育種更順利，可利用熱休克、秋水仙素或氮氣處理，以增加2n花粉的產生。
4、另外在利用上要注意花粉的新鮮度 (開花一天後的花粉發芽率<5%)，才能使2n花粉順利發芽。

55. Conclusion Utilization of 2n-pollen in Lilium:
2n-Pollen from both OA hybrids showed fairly good germination when fresh pollen was used (40% in and 100% in ).
Utilization of 2n-pollen in Lilium:
It is important to note that the use of fresh 2n-pollen grains can ensure success for crosses.
1、 透過2N花粉的利用，使具有不稔性的百合，再度成為有利用價值的親本。
2、2N花粉配子具有種間雜交染色體互換重組的能力，可以產生更多的遺傳變異，增加遺傳岐異度，對育種是有利的。
3、但並目前仍儘有少數的雜交種可以產生較多的2n-花粉，為使育種更順利，可利用熱休克、秋水仙素或氮氣處理，以增加2n花粉的產生。
4、另外在利用上要注意花粉的新鮮度 (開花一天後的花粉發芽率<5%)，才能使2n花粉順利發芽。

56. Thanks
for your attention