Plant Cell Reports
Plant Cell Reports (1988) 7: 127-129
© Springer-Verlag 1988
Substitution analysis of callus induction and plant regeneration from anther culture in wheat (Triticum aestivum L.) l~va Szakfics ~, G6za Kovfics ~, Jfinos Pauk 2, and Befita Barnabfis ~ Agricultural Research Institute of the Hungarian Academy of Sciences, Martonvfisfir, H-2462, Pf. 19, Hungary Cereal Research Institute, Szeged, H-6701, Pf. 391, Hungary Received September 16, 1987 / Revised version received December 15, 1987 - Communicated by H. L6rz
Abstract The oenetie determinati.oH of callus ind,,orion, total plant regener_ation and green plant re qeneration from aotiler cu]t~re ~,,,ere studied using a "Chinese Spring"/'!Cheyenne" s~bstit~tion series. A]l the three characteristics ~,~ere found to be polygeoical]y determined, but their inheritance was independent f.rom one another. The 7A and l~ chromosomes had a considerable elf.cot on ea]lus induction, In the case of. total plant regeneration the most influential chromosome ,,as the 3A ~hi]e the 2D chromosome shorted a def.inite influence on green plant reoeneration. The interaction bet~-~een the oenetic background of. the recipient plant and the substituted chromosome plays an important role in the manifestation of the studied f.eatures.
Introduction Since haploid p!ants ,.,ere first identified (elakes]ee et al. 1922) and produced in vitro from anther culture (Guha and Mahesh~,ari 19~4) the importance of. this technique f.or practical plant breeding has increased (Griff.ing 1975). Routine application of anther cultures in plant breeding ~,ill not be possible ,,ntil large numbers of. plants can be produced. Experience relating to anther cultures gives rise to the conclusion that microspore callus induction considerably depends on the gro~ing conditions of. pollen donor plants (Wang and Che.n 1980, Lazar et al. 1983), the in vitro culture conditions (Jing et al. 1982), the developmental stage of. microspores (Ouyang eta].. 1983) and the genotype of. the donor plant (Bullock et al. 1982). Anther culture as an experimental system might provide a lot of information in relation to the follo!~!ino problems: 1. Studies on the gro!~ing conditions of. anther donor plants 2. Increasing the ef.ficiency of. in vitro techniques 3. Revealing the genetic determination of. in vitro processes. Several authors have reported the genetic determination of the oro,,,th of hap~:oid and
Offprint requests to." E. Szak~tcs
diploid calli in ~,.,heat. The frequency of. cal1L,s induction from anthers sho!,led sJ.onJ_fica,lt variation bet,,reen aneuploid lines of "Chinese Spring" t~!heat varieties (Shimada and Making 1975). Similar results ,,ere obtained f.rom tissue cultures arising from immature embryos of a "Chinese Sprino"/"Chayenne" substitutiorl series (Galiba et al. ].986). Matthias and Fukui (1986) reported the effect of specif.ic chromosome and cytoplasm substitl]tions on the tissue culture responses of_ calli of four lines of. "Chinese Spring". The aim of the present study ,~as to carry o~,t an experiment to investigate the elf.cots of. specif.ic chromosomes on the in vitro anther culture process. Materials
and methods
Plant material The "Chinese Spring"/"Cheyenne" substitution series ,~as developed at the University of Nebraska~ USA. Each line of. the series consists of. a "Chinese Spring" background of. 21 chromosome pairs ~,ith a different "Cheyenne" chromosome pair substituted in turn f.or the corresponding homologous pair of the "Chinese Spring" complement. Spikes ~Jere collected f.rom plants raised in the f.ield. Anther culture Before excision of anthers spikes ~,sere sterilized ,~ith O.iZ HgCI solution and ~,4ashed with distilled !later. The tissue cultures ~Jere induced from anthers containino microspores at uninucleate stage. On the average 1800 anthers ~,ere incubated per line ~,Jith 250-300 anthers in each petri dish. P-2 medium containing 1.5 mg/1 2,4-0 ~as used for microspore callus induction (Chuang et a].. 1978). The cultures ,~,~ere incubated ~or 40 days at a temperature of. 29°C in the dark. Plants !,,~ere regenerated on 190-2 medium (He and Ouyang 1984) ~,~ithout 2,4-D at a temperature of. 26°C ~.,ith a 16-hour illumination. Callus induction ,~las calcu]ated on the basis of the number of inoculated anthers. Plant regeneration ~'!as counted on the basis of calli cultured Analysis of variance was carried out on the data for comparison.
128
Results Callus
Table 2. Plant regeneration from anther calli of "Chinese Spring"/"Cheyenne" substitution liformation
nes
The res.].ts are summarised in Table i. Table ]., Ca]_lus induction from the anthers of "Chinese Spring"/"Cheyenne" substitution lines Substitutiofl lines
Number of Induced ca]li anthers number %,
Deviation from "Chzn " ~.s,R Sp..J.no r" . "
CS Ch IA IB
2276 4117 1228 871
409 807 ],80 2
18.0 ].9.6 ].4.7 0.2
Z N.O. . . . + i. 6 - 3.3 - 1 7 . 8 ××
ID 2A 28 20
154,2 1700 970 2423
41 24 32 13].
2.7 I.~ 3.3 5.4
-15.3~ x -16.6 ×x -14.7 xx -12.6 x
3A
1608
127
7.9
-iO.l ×
38 30 ZBA
2200 2162 2~.i0
81 91 367
3.7 4.2 15.2
-14.3 x -13.8 x - 2•8
48
829
122
- 3.3
~O 5A 58 5D 6A
1278 2688 ].z!.92 4232 1138 1306
63 2].8 31 199 , 41 ].63
14.7 ~..9
1898 1400
I18 O 196
68 60 7A 78 70
Iota].
].284 1150 /~22].2
60 3883
8.-I 2.1 11.on
3.6 12.5
-13.1 x - 9.9 -15.9 xx - 6.2 -14.4 xX -:5.5
6.2 O.O
-].1.8" -18.0 ×
15.3 5.2 9.3
- 2.7 -12.8 x -
x Sionificantat 5% level, xx Significant at i% level From the point of view of callus induction no differences could be observed between the "Chinese Spring" (recipient) and "Cheyenne" (donor) wheat varieties• However the callus induction abilities of the s u b s t i t u t i o n lines were considerably lower than those of the two varieties. The 7A and 1B s u b s t i t u t i o n s showed a p r o m i n e n t l y lo~ callus induction frequency. In the case of 7A there was no callus induction and the induction frequency of 18 ~as so low that it can be c o n s i d e r e d p r a c t i c a l l y zero. Besides the two most influential c h r o m o s o m e s the 1D, 2A, 28, 58 and 6A c h r o m o s o m e s also c o n s i d e r a b l y decreased the callus indflction ability. The 2B, 3A, 3D 1 40, 60 and 78 chromosomes had lesser effects• The other chromosomes had no effect on callus induction ability. Haploid
plant
regeneration
The 7A and 1B s u b s t i t u t i o n lines ~lere omitted from this study as they produced no calli. Most of the r e g e n e r a t e d plants were albinos. The results were divided into total plant r e g e n e r a t i o n and green plant r e g e n e r a t i o n (Table 2).
Substitution lines
Number of calli cu]_tured
CS Ch IA 1B ID 2A 28
Plant regeneration frequency Total number Green plants of plants % deviation % deviation from "CS" from "CS"
409 807 180 2 41 24
15.1 6.6 10.5 . 9.5 7=7
0.~ - 8. x - 4.6 . . - 5.6 - 7.4
4.1 2.0 2.5
0]~ x - 2 - 1.6
4.8
+ 0.7 - 4:1'!".i x×
.
32
].0•3
- a 8
O.O O.O
2D 3A
131 127
12.5 30.2
- 2.6 +15.1×x
5.9 3.5
+ 1.8 × - 0.6
38 30 4A
81 91 367
3. I~ 3•5 17.7
- i i • 7x -11.6 x + 2.6
O. 0 0.0
- l,. i X× - 4.1 xx - 1•8,"]
48
122
8.9
- 6.2x×
40 5A 5B 50 6A 6B 6D 7A 78
63 218
1.7 22.1
31
23.3
499 41 163 118 . 196
12,4 8.3 7.3 10.2 . 7.6
-13.4 + 6.9 + 8.2 )" - 2.7 - 6.8 -7.8 - 4.9 . . - 7.5
7D
60
15.2
+ 0.i
2.3 0.9 O.O 1.5 O. O ]_.4 O.O 2.7 1.7
2.3 1.7
4
3 2 -
4.• l×X 2.6 )< z,~.1 ''× 2.7 x 4.1 ×x 1.4 2.4 ×
- 1.7
- 2.3 x
x Significant at 5Z level, x× SionifiOant at 1% level C o n c e n r i n g the total plant r e g e n e r a t i o n the "Chinese Spring" and "Cheyenne" wheat varieties differ from each other. Far fewer plantlets could be r e g e n e r a t e d from "Cheyenne" than from "Chinese Spring" Most of the lines also had lower r e g e n e r a t i o n capacity than the recipient variety but there were exceptions. The 3A c h r o m o s o m e s i g n i f i c a n t l y increased the plant r e g e n e r a t i o n frequency and the 58 chromosome had a similar, but lesser, effect. The 40, 38 and 3D c h r o m o s o m e s strongly decreased r e g e n e r a t i o n capacity. The other c h r o m o s o m e s seemed to have no influence on the r e g e n e r a t i on process. Data on green plant r e g e n e r a t i o n are very different from those of total plant reg e n e r a t i o n because of the high frequency of albino plantlets. More green plantlets could be r e g e n e r a t e d from "Cheyenne". It is an interesting p h e n o m e n o n that seven lines did not respond in green plant r e g e n e r a t i o n at all but there were two lines which showed increased response. C h r o m o s o m e 2D had the strongest effect and lO also had a slight effect. All the other c h r o m o s o m e s generally decreased the green plant r e g e n e r a t i o n capacity. Discussion From the 42,212 anthers cultured, 3,802 (9.3%) calli l~ere induced and 82 (2.2%) green plants r e g e n e r a t e d from them. According to the data (Table 3) the three studied features can be supposed to be p o l y g e n i c a l l y determined.
129 Table
3 Effect of_ s u b s t J . t u t e d chromosomes on the frequencies of callus formation, plant regeneration and green plant production
in a direct genetic way and to produce genotypes which contain gene combinations advantageous for anther culture. References
Substitution ]_ines 'CS Ch l& ].8 iO 2A 28 20 3A
3,3
CallJs formafinn St Ns NS -xx -xx -xx -xx - x - x - X
St -xx Ns Nt Ns Ns Ns Ns
Green plant regeneration St - x Ns Nt Ns -xx -xx + x
+xx
Ns
- x
-xx
-
--xy - x - x
30 4A 4,r4 t,q)
-
5&
Ns -xx Ns
Ns Ns Ns
- x -'~x - x
6A .GB
-xx
Ns Ns
-
Ns
40
- x -xx Ns - x
Ns Nt Ns Ns
5,3 513
7A 78 713
x Ns Ns
Plant regeneration
-
x
x Ns Ns
-xx
-xx
x
Ns - ), Nt Ns - x
Ns - not significant~ x - significant at 5% level, xx - significant at 1% level, N-~ - not tested. + and - - tendency d%_ the effect as compared to "CS", St - "CS" standard. 3esides several chromosomes which reduced to different extents the haploid callus induction capacity, in the presence of chromosomes 7A and 1B there were no calli induced. The 7A and 18 chromosomes probably carry major genes influencing this process. The fact that there ,,Jas no difference bet,,~een the two varieties in callus induction ability indicates that t h e interaction between the genetic background Of the recipient plant and the substituted chromosome might be very important. Hypothetically a given chromosome can have different effects in different genetic backgrounds. It w o u l d be necessary t o study the 7A and 1B chromosomes in other wheat varieties. If there is a multiplex allelic variation on callus induction then it would, be possible to identif.y chromosomes 7A and ].8 and manipulate ,~Jith them for the purpose of increasing callus induction in different varieties. In respect of total and green plant regeneration ability the two studied varieties differed s i g n i f i c a n t l y from each other. We had supposed that all of the chromosomes would reduce the plant regeneration capacity in comparison with the recipient variety. However it was folmd that chromosome 3A increased the frequency of total plant regeneration and 28 increased the green plant regeneration capacit y. The data indicate that the three different processes examined might be under different genetic regu]_ation and can be inherited independently as it was also suggested by Lazar et al. (1984) and Oeaton et al. (1987) on the basis of their results from anther c[llture of ,;,heat This could possibly give an opportunity to solve the problem of' albinism
Blakeslee AF, Belling J Farnham ME, BerBer AO (1922) A haploid mutant in the .iimson ~leed, gatura stramonium. Science 55: 646-647. Bullock WO, Baenzinger PS, Schaeffer GW, 8offing PJ (1982) Anther culture of wheat (Triticum aestivum L.) Fl's and their reciprocal crosses. Theor Appl Genet 6 2 : 1 5 5 - 1 5 9 Chuang CC, Ouyang TW Chia H Chou SM Chin CK (1978) A set of potato media for wheat anther cult,re. In Proc Symp Plant Tissue Culture, Science Press Peking pp 51-56 Oeaton WR, Metz SG, Armstrong TA, Mascia PN (1987) Genetic analysis of the anther-culture response of th#ee spring wheat crosses. Theor Appl Genet 74: 334-338. Galiba 8, Kov~cs G, Sutka J (1986) Substitution analysis of plant regeneration from callus culture in wheat. Plant Breeding 97: 26].-263 6rif#.ing B (].975) Efficiency changes due to use of_ doubled-haploids in recurrent selection methods. Theor Appl Genet 1~6: 367-386 Guha S, Maheshwari SC (].96@) In vitro production of embryo from anther of gatura. Nature 204: 497 He OG, Ouyang 3W (1984) Callus and plantlet formation from cultured wheat anthers at different developmental stages. Plant Sol Let 33: 71-79 Jing JK, Xi ZY, Hu 14 (1982) Effects of high temperature and physiological condition of donor plants on induction of pollen derived plants in t,#heat. Ann Rep Inst Genet Acad Sinic Peking 1 9 8 1 : 6 7 - 6 8 Lazar MD, Baenziger PS, Schaeffer GW (1984) Combining abilities and heritability of callus formation and plantlet regeneration in wheat (Triticum aestivum L.) anther cultures. Theor Appl Genet 6 8 : 1 3 1 - 1 3 4 Lazar MB, Collins GB, Vian WE (1983) Genetic and environmental effectsson the gre,,~,th of differentiation of wheat somatic cell cultures. J Hered 74: 353-357 Lazar MO, Chen THH, Scoles 8J, Kartha KK (198z~) Immature embryo and anther culture of chromosome addition lines of rye in Chinese Spring ,,Jheat. Plant Science 5 1 : 7 7 - 8 1 Mathias RJ, Fukui K (1986) The effect of specific chromosome and cytoplasm substitutions on the tissue culture response of wheat (Tr~ticum aestiwJm) callus. Theor Appl Genet 7 1 : 7 9 7 - 8 0 0 guyang TW, Chuang CO, Tseng CC (1973) Induction of pollen plants from anther of Triticure aestiwJm L. cultured in vitro. Sci Sinice 16: 79-95 Shimada T, Making T (1975) In vitro c,lture of wheat. III. A,nther culture of the A genom aneuploids in common wheat. Theor Appl 6enet 46:t~07-418 Wang P, Chen YR (].980) Effects of gro~,Jth conditions of anther-donor plants on the production of pollen plants in wheat anther culture. Aota Genet Sinic 7:6z,~-71
Plant Cell Reports (1988) 7: 127-129
© Springer-Verlag 1988
Substitution analysis of callus induction and plant regeneration from anther culture in wheat (Triticum aestivum L.) l~va Szakfics ~, G6za Kovfics ~, Jfinos Pauk 2, and Befita Barnabfis ~ Agricultural Research Institute of the Hungarian Academy of Sciences, Martonvfisfir, H-2462, Pf. 19, Hungary Cereal Research Institute, Szeged, H-6701, Pf. 391, Hungary Received September 16, 1987 / Revised version received December 15, 1987 - Communicated by H. L6rz
Abstract The oenetie determinati.oH of callus ind,,orion, total plant regener_ation and green plant re qeneration from aotiler cu]t~re ~,,,ere studied using a "Chinese Spring"/'!Cheyenne" s~bstit~tion series. A]l the three characteristics ~,~ere found to be polygeoical]y determined, but their inheritance was independent f.rom one another. The 7A and l~ chromosomes had a considerable elf.cot on ea]lus induction, In the case of. total plant regeneration the most influential chromosome ,,as the 3A ~hi]e the 2D chromosome shorted a def.inite influence on green plant reoeneration. The interaction bet~-~een the oenetic background of. the recipient plant and the substituted chromosome plays an important role in the manifestation of the studied f.eatures.
Introduction Since haploid p!ants ,.,ere first identified (elakes]ee et al. 1922) and produced in vitro from anther culture (Guha and Mahesh~,ari 19~4) the importance of. this technique f.or practical plant breeding has increased (Griff.ing 1975). Routine application of anther cultures in plant breeding ~,ill not be possible ,,ntil large numbers of. plants can be produced. Experience relating to anther cultures gives rise to the conclusion that microspore callus induction considerably depends on the gro~ing conditions of. pollen donor plants (Wang and Che.n 1980, Lazar et al. 1983), the in vitro culture conditions (Jing et al. 1982), the developmental stage of. microspores (Ouyang eta].. 1983) and the genotype of. the donor plant (Bullock et al. 1982). Anther culture as an experimental system might provide a lot of information in relation to the follo!~!ino problems: 1. Studies on the gro!~ing conditions of. anther donor plants 2. Increasing the ef.ficiency of. in vitro techniques 3. Revealing the genetic determination of. in vitro processes. Several authors have reported the genetic determination of the oro,,,th of hap~:oid and
Offprint requests to." E. Szak~tcs
diploid calli in ~,.,heat. The frequency of. cal1L,s induction from anthers sho!,led sJ.onJ_fica,lt variation bet,,reen aneuploid lines of "Chinese Spring" t~!heat varieties (Shimada and Making 1975). Similar results ,,ere obtained f.rom tissue cultures arising from immature embryos of a "Chinese Sprino"/"Chayenne" substitutiorl series (Galiba et al. ].986). Matthias and Fukui (1986) reported the effect of specif.ic chromosome and cytoplasm substitl]tions on the tissue culture responses of_ calli of four lines of. "Chinese Spring". The aim of the present study ,~as to carry o~,t an experiment to investigate the elf.cots of. specif.ic chromosomes on the in vitro anther culture process. Materials
and methods
Plant material The "Chinese Spring"/"Cheyenne" substitution series ,~as developed at the University of Nebraska~ USA. Each line of. the series consists of. a "Chinese Spring" background of. 21 chromosome pairs ~,ith a different "Cheyenne" chromosome pair substituted in turn f.or the corresponding homologous pair of the "Chinese Spring" complement. Spikes ~Jere collected f.rom plants raised in the f.ield. Anther culture Before excision of anthers spikes ~,sere sterilized ,~ith O.iZ HgCI solution and ~,4ashed with distilled !later. The tissue cultures ~Jere induced from anthers containino microspores at uninucleate stage. On the average 1800 anthers ~,ere incubated per line ~,Jith 250-300 anthers in each petri dish. P-2 medium containing 1.5 mg/1 2,4-0 ~as used for microspore callus induction (Chuang et a].. 1978). The cultures ,~,~ere incubated ~or 40 days at a temperature of. 29°C in the dark. Plants !,,~ere regenerated on 190-2 medium (He and Ouyang 1984) ~,~ithout 2,4-D at a temperature of. 26°C ~.,ith a 16-hour illumination. Callus induction ,~las calcu]ated on the basis of the number of inoculated anthers. Plant regeneration ~'!as counted on the basis of calli cultured Analysis of variance was carried out on the data for comparison.
128
Results Callus
Table 2. Plant regeneration from anther calli of "Chinese Spring"/"Cheyenne" substitution liformation
nes
The res.].ts are summarised in Table i. Table ]., Ca]_lus induction from the anthers of "Chinese Spring"/"Cheyenne" substitution lines Substitutiofl lines
Number of Induced ca]li anthers number %,
Deviation from "Chzn " ~.s,R Sp..J.no r" . "
CS Ch IA IB
2276 4117 1228 871
409 807 ],80 2
18.0 ].9.6 ].4.7 0.2
Z N.O. . . . + i. 6 - 3.3 - 1 7 . 8 ××
ID 2A 28 20
154,2 1700 970 2423
41 24 32 13].
2.7 I.~ 3.3 5.4
-15.3~ x -16.6 ×x -14.7 xx -12.6 x
3A
1608
127
7.9
-iO.l ×
38 30 ZBA
2200 2162 2~.i0
81 91 367
3.7 4.2 15.2
-14.3 x -13.8 x - 2•8
48
829
122
- 3.3
~O 5A 58 5D 6A
1278 2688 ].z!.92 4232 1138 1306
63 2].8 31 199 , 41 ].63
14.7 ~..9
1898 1400
I18 O 196
68 60 7A 78 70
Iota].
].284 1150 /~22].2
60 3883
8.-I 2.1 11.on
3.6 12.5
-13.1 x - 9.9 -15.9 xx - 6.2 -14.4 xX -:5.5
6.2 O.O
-].1.8" -18.0 ×
15.3 5.2 9.3
- 2.7 -12.8 x -
x Sionificantat 5% level, xx Significant at i% level From the point of view of callus induction no differences could be observed between the "Chinese Spring" (recipient) and "Cheyenne" (donor) wheat varieties• However the callus induction abilities of the s u b s t i t u t i o n lines were considerably lower than those of the two varieties. The 7A and 1B s u b s t i t u t i o n s showed a p r o m i n e n t l y lo~ callus induction frequency. In the case of 7A there was no callus induction and the induction frequency of 18 ~as so low that it can be c o n s i d e r e d p r a c t i c a l l y zero. Besides the two most influential c h r o m o s o m e s the 1D, 2A, 28, 58 and 6A c h r o m o s o m e s also c o n s i d e r a b l y decreased the callus indflction ability. The 2B, 3A, 3D 1 40, 60 and 78 chromosomes had lesser effects• The other chromosomes had no effect on callus induction ability. Haploid
plant
regeneration
The 7A and 1B s u b s t i t u t i o n lines ~lere omitted from this study as they produced no calli. Most of the r e g e n e r a t e d plants were albinos. The results were divided into total plant r e g e n e r a t i o n and green plant r e g e n e r a t i o n (Table 2).
Substitution lines
Number of calli cu]_tured
CS Ch IA 1B ID 2A 28
Plant regeneration frequency Total number Green plants of plants % deviation % deviation from "CS" from "CS"
409 807 180 2 41 24
15.1 6.6 10.5 . 9.5 7=7
0.~ - 8. x - 4.6 . . - 5.6 - 7.4
4.1 2.0 2.5
0]~ x - 2 - 1.6
4.8
+ 0.7 - 4:1'!".i x×
.
32
].0•3
- a 8
O.O O.O
2D 3A
131 127
12.5 30.2
- 2.6 +15.1×x
5.9 3.5
+ 1.8 × - 0.6
38 30 4A
81 91 367
3. I~ 3•5 17.7
- i i • 7x -11.6 x + 2.6
O. 0 0.0
- l,. i X× - 4.1 xx - 1•8,"]
48
122
8.9
- 6.2x×
40 5A 5B 50 6A 6B 6D 7A 78
63 218
1.7 22.1
31
23.3
499 41 163 118 . 196
12,4 8.3 7.3 10.2 . 7.6
-13.4 + 6.9 + 8.2 )" - 2.7 - 6.8 -7.8 - 4.9 . . - 7.5
7D
60
15.2
+ 0.i
2.3 0.9 O.O 1.5 O. O ]_.4 O.O 2.7 1.7
2.3 1.7
4
3 2 -
4.• l×X 2.6 )< z,~.1 ''× 2.7 x 4.1 ×x 1.4 2.4 ×
- 1.7
- 2.3 x
x Significant at 5Z level, x× SionifiOant at 1% level C o n c e n r i n g the total plant r e g e n e r a t i o n the "Chinese Spring" and "Cheyenne" wheat varieties differ from each other. Far fewer plantlets could be r e g e n e r a t e d from "Cheyenne" than from "Chinese Spring" Most of the lines also had lower r e g e n e r a t i o n capacity than the recipient variety but there were exceptions. The 3A c h r o m o s o m e s i g n i f i c a n t l y increased the plant r e g e n e r a t i o n frequency and the 58 chromosome had a similar, but lesser, effect. The 40, 38 and 3D c h r o m o s o m e s strongly decreased r e g e n e r a t i o n capacity. The other c h r o m o s o m e s seemed to have no influence on the r e g e n e r a t i on process. Data on green plant r e g e n e r a t i o n are very different from those of total plant reg e n e r a t i o n because of the high frequency of albino plantlets. More green plantlets could be r e g e n e r a t e d from "Cheyenne". It is an interesting p h e n o m e n o n that seven lines did not respond in green plant r e g e n e r a t i o n at all but there were two lines which showed increased response. C h r o m o s o m e 2D had the strongest effect and lO also had a slight effect. All the other c h r o m o s o m e s generally decreased the green plant r e g e n e r a t i o n capacity. Discussion From the 42,212 anthers cultured, 3,802 (9.3%) calli l~ere induced and 82 (2.2%) green plants r e g e n e r a t e d from them. According to the data (Table 3) the three studied features can be supposed to be p o l y g e n i c a l l y determined.
129 Table
3 Effect of_ s u b s t J . t u t e d chromosomes on the frequencies of callus formation, plant regeneration and green plant production
in a direct genetic way and to produce genotypes which contain gene combinations advantageous for anther culture. References
Substitution ]_ines 'CS Ch l& ].8 iO 2A 28 20 3A
3,3
CallJs formafinn St Ns NS -xx -xx -xx -xx - x - x - X
St -xx Ns Nt Ns Ns Ns Ns
Green plant regeneration St - x Ns Nt Ns -xx -xx + x
+xx
Ns
- x
-xx
-
--xy - x - x
30 4A 4,r4 t,q)
-
5&
Ns -xx Ns
Ns Ns Ns
- x -'~x - x
6A .GB
-xx
Ns Ns
-
Ns
40
- x -xx Ns - x
Ns Nt Ns Ns
5,3 513
7A 78 713
x Ns Ns
Plant regeneration
-
x
x Ns Ns
-xx
-xx
x
Ns - ), Nt Ns - x
Ns - not significant~ x - significant at 5% level, xx - significant at 1% level, N-~ - not tested. + and - - tendency d%_ the effect as compared to "CS", St - "CS" standard. 3esides several chromosomes which reduced to different extents the haploid callus induction capacity, in the presence of chromosomes 7A and 1B there were no calli induced. The 7A and 18 chromosomes probably carry major genes influencing this process. The fact that there ,,Jas no difference bet,,~een the two varieties in callus induction ability indicates that t h e interaction between the genetic background Of the recipient plant and the substituted chromosome might be very important. Hypothetically a given chromosome can have different effects in different genetic backgrounds. It w o u l d be necessary t o study the 7A and 1B chromosomes in other wheat varieties. If there is a multiplex allelic variation on callus induction then it would, be possible to identif.y chromosomes 7A and ].8 and manipulate ,~Jith them for the purpose of increasing callus induction in different varieties. In respect of total and green plant regeneration ability the two studied varieties differed s i g n i f i c a n t l y from each other. We had supposed that all of the chromosomes would reduce the plant regeneration capacity in comparison with the recipient variety. However it was folmd that chromosome 3A increased the frequency of total plant regeneration and 28 increased the green plant regeneration capacit y. The data indicate that the three different processes examined might be under different genetic regu]_ation and can be inherited independently as it was also suggested by Lazar et al. (1984) and Oeaton et al. (1987) on the basis of their results from anther c[llture of ,;,heat This could possibly give an opportunity to solve the problem of' albinism
Blakeslee AF, Belling J Farnham ME, BerBer AO (1922) A haploid mutant in the .iimson ~leed, gatura stramonium. Science 55: 646-647. Bullock WO, Baenzinger PS, Schaeffer GW, 8offing PJ (1982) Anther culture of wheat (Triticum aestivum L.) Fl's and their reciprocal crosses. Theor Appl Genet 6 2 : 1 5 5 - 1 5 9 Chuang CC, Ouyang TW Chia H Chou SM Chin CK (1978) A set of potato media for wheat anther cult,re. In Proc Symp Plant Tissue Culture, Science Press Peking pp 51-56 Oeaton WR, Metz SG, Armstrong TA, Mascia PN (1987) Genetic analysis of the anther-culture response of th#ee spring wheat crosses. Theor Appl Genet 74: 334-338. Galiba 8, Kov~cs G, Sutka J (1986) Substitution analysis of plant regeneration from callus culture in wheat. Plant Breeding 97: 26].-263 6rif#.ing B (].975) Efficiency changes due to use of_ doubled-haploids in recurrent selection methods. Theor Appl Genet 1~6: 367-386 Guha S, Maheshwari SC (].96@) In vitro production of embryo from anther of gatura. Nature 204: 497 He OG, Ouyang 3W (1984) Callus and plantlet formation from cultured wheat anthers at different developmental stages. Plant Sol Let 33: 71-79 Jing JK, Xi ZY, Hu 14 (1982) Effects of high temperature and physiological condition of donor plants on induction of pollen derived plants in t,#heat. Ann Rep Inst Genet Acad Sinic Peking 1 9 8 1 : 6 7 - 6 8 Lazar MD, Baenziger PS, Schaeffer GW (1984) Combining abilities and heritability of callus formation and plantlet regeneration in wheat (Triticum aestivum L.) anther cultures. Theor Appl Genet 6 8 : 1 3 1 - 1 3 4 Lazar MB, Collins GB, Vian WE (1983) Genetic and environmental effectsson the gre,,~,th of differentiation of wheat somatic cell cultures. J Hered 74: 353-357 Lazar MO, Chen THH, Scoles 8J, Kartha KK (198z~) Immature embryo and anther culture of chromosome addition lines of rye in Chinese Spring ,,Jheat. Plant Science 5 1 : 7 7 - 8 1 Mathias RJ, Fukui K (1986) The effect of specific chromosome and cytoplasm substitutions on the tissue culture response of wheat (Tr~ticum aestiwJm) callus. Theor Appl Genet 7 1 : 7 9 7 - 8 0 0 guyang TW, Chuang CO, Tseng CC (1973) Induction of pollen plants from anther of Triticure aestiwJm L. cultured in vitro. Sci Sinice 16: 79-95 Shimada T, Making T (1975) In vitro c,lture of wheat. III. A,nther culture of the A genom aneuploids in common wheat. Theor Appl 6enet 46:t~07-418 Wang P, Chen YR (].980) Effects of gro~,Jth conditions of anther-donor plants on the production of pollen plants in wheat anther culture. Aota Genet Sinic 7:6z,~-71
