Copyright is owned by the Author of the thesis. Permission is given for
a copy to be downloaded by an individual for the purpose of research and
private study only. The thesis may not be reproduced elsewhere without
the permission of the Author.
PROPAGATION OF ACTINIDIA CHINENSIS (PLANCH.) BY STEM AND ROOT CUTTING
A Thesis Submitted in partial lulfilment of the requirements for the degree of
Master in Horticultural Science at Massey University
SIM BOON LIANG
1979
(ii)
ABSTRACT
Basal wounding, bottom heat, light with IBA treatments were found to be beneficial for rooting of
_Actinidia chinensis (Planch.). IBA treatment was effective only when there was a high natural ability to initiate root in Summer and Spring.
Seasonal fluctuations in rooting ability was pronounced. This seasonal variation seems to be related to the levels of endogenous IAA, ABA and cofactor 2. No correlation between root initiation and bud activity or
IAN level was established.
IAA seems to be the fundamental physiological promoter of adventitous root formation. IBA plays only a supporting role in promoting root formation, by protecting the endogenous IAA level in the cutting base.
Leaf tissue is an important factor for rooting to be successful. The role of leaf tissue is not just to
produce auxin or synthesize nutrients but rather some unknown factor in the leaf can produce a synergistic interaction with auxin in root formation processes.
Root cuttings of Abbott variety were sequentially harvested and planted over a period from late Autumn
(1.4.77) until mid Summer (8.1.78). Root cuttings of different thickness and length were compared to evaluate their effect on regeneration. The effect of various growt regulators was investigated too.
Root diameters of 0.5 - 1.5 cm. out performed that of the thinner or thicker ones. Shorter cuttings (5 cm) of equivalent total length were found to be more productive than a single long cutting (15cm). Strong polarity was observed with shoots only arising from the proximal end of the cutting.
Regenerative capacity was highest in late Autumn and Winter and lowest in Summer. This seasonal fluctuation
(iii) can be altered by exogenous application of growth
regulators. IBA suppressed shoot regeneration, whereas cytokinin and sucrose promoted it, while GA3 did not have any significant effect.
For commercial use, the practical and economic aspects of this techique require further investigation.
IAA IAN IBA
ABA
GA3
GA
BAP
DNA RNA
PPM
p
~
IX
~ll
Glossary of Abbreviations
=
::
=
:a
=
=
:z
=
=
=
=
=
=
=
indoleacetic acid indoleacetic nitrile indolebutyric acid abscisic acid
gibberellic acid gibberellin
benzylaminoptlrine
deoxyribonucleic acid ribonucleic acid
parts per million probability
0 .196 0.0596 0.00196
(iv)
(v)
ACKNOWLEDGEMENTS
The author would like to thank Professor
J.A. Veale, Mr. G.S. Lawes and Dr. D.J. Woolley for their guidance and assistance in their capacity as supervisors.
My sincere thanks to Mr. G. Vander Mespel of Horticultural Research Centre in Levin for supplying cutting materials.
I would like to express my gratitude to A. Watson and D. Anderson for their technical assistance.
C O
=========•=====
N T E N T SChapter
1 Review of Literature 1.1 Introduction
1.2 Propagation of Stem Cuttings
1.2.1 Anatomical Development of Roots Cuttings
1.2.1.1 Mechanical hinderance 1.2.2 Environmental Factors Affecting
Rooting Ability of Cutting 1 .2.2.1 Seasonal Variation 1.2.2.2
1.2.2.3
Photo period
Other Light Effect
1.2.2.4 Physical Envirorunent for Rooting 1.2.3 Physiological Aspects
1.2.3.1 Juvenility
1.2.3.2 The Effects of Leaves 1.2.3.3 The Effects of Buds
1.2.3.4 The Effects of Flowering Buds 1.2.4 Hormonal Basis of Rooting
1.2.4.1 1.2.4.2 1.2.4.3 1.2.4.4 1.2.4.5
Endogenous Auxins Oytokinin
Gibberellins and Rooting Abscisic Acid
Ethylene 1.2.5 Role of Cofactors 1.2.6 Nutrition
1.2.6.1 Carbohydrate 1.2.6.2 Nitrogen
1.2.6.3 Other Nutritions
1.2.7 Type of Wood Selected for Cuttings 1 .3 Root Cuttings
1 1 1
2 4
5 5 9 11 13 15 15 17 20 22 23 24
28
29 35 37 38 40 41 43 44 45
46
Chapter
2 Rationale of the Pre sent Work
3 General Material s and Methods 3.1 Plant Materials
3.2 Collectjon and Preparation of Cutting Materials
3,3 Treatment and Planting of Cutting 3.3.1
3.3.2 3.3.3
Wounding and Rooting Condition Indole Butyric Acid Treatment Application of Indole-Acetic Acid 3.4 Plast ic Tent
'3.5 Sampling of Material for !Iorrnone Extraction
'3.1; Extraction of PJant Mater.ial for Hormone Annlysis
·-s.6 .1 Initial Extract ion for Auxin and Inhibitors
3.6.2 Extraction of PJant Material for Root ing Cofactors
7
• 7 C~1rcmatography of Extr'tcts for Hormone Analysis
3. 7. 1 Paper Chromatogrc111hy of Auxin and Inhibitors
3.7.2 Paper Chromatography of Rooting Cofactors
3.8 Bioassay Procedures
3.8.1 Triticum Coleoptile Bioassay 3.8.2 Avena Coleoptile Bioassay
3.8.3 Mung Bean Root Initiation Bioassay 3.9 Root Cuttings
49 50 50 50 50 50 51 52 52
53 53 53
57 57 57 58 58 58 59 64 66
Chapter
4 Propagation of Actinidia chinensis by Stem Cuttings
4.1 The Effect of Wounding on H.ooting Actinidia chinensis
4.2 Varietal and Sexual Difference in the Pro-pagation of Actinidia e}1incnsis 4. 3 Propagation of Leafy r:11: t i r,r :· u.1der
Polythene-Tent
4 .4 The Effect of Bxop-enou., Tl" ,u1 Rooting of Actinidia chinensi s
4.4.1 The Influence 8t' T// n·l Bar.ml Temperature on t.11, tlc•otj np; of Actinidia chine •:;; .·
4.4.2 The Effect of V r:ous Concentration of IBA on Root h1·r1'llf'!'i1tion of
67 67
70 72
76
Actinidia chinen:::;is 79
5
r ;.
5 Sec1sonal Chanres in Hoot Ini ti8.tion of Actinidia chinensis4. ,-; The Relationship Between dud Dormancy and Root Initiation
·1 . 7 Seasonal Changes of Endogenous Growth Regulators of Actinidln chincnsis
.t . r3 E:xcJ.mination of the Rol e of I/IA and IBA in Root Formation of Bruno Cuttings 4.9 Effects of Leaves
Propagatin,£? Actinidia chinensis by Root Cutting
5. 1 Introduction
5.2 The Shoot Regeneration Potential of Roots of Different Diameter
5.3 Root Length and Polarity 5.4 Seasonal Effects
5.5 Applied Growth Regulators
89
95 101 113 130
137 137 137 140 145 147
Chapter
6 Gener al Discussion and Conclusion 6.1 Stem Cuttings
6.2 Root Cuttings
7 Anpend.ices
Appendix 1 Standard Buffer Solution
Appendix 2 ANOVA for Stem Cutting Experiments Appendix 3 ANOVA for Root Cutting Experiments
8 8j bliography
Page
152 152 161
165 165 166 174
177
Fig. 3. €1.1
Fi.f. 3.8. i
Fir. 3.8.2
Fir-. 3.8.3
·r
c1 b10 /4 • 1 • 1rr ~, b J e 4 • 2 • 1
'I ; 1 \J 1 ,, •I • ':5 • 1
LIST OF FIGURES AND TABLES
= ===== ==== === ===--===~== ======
Summary of hor:none extraction procedure
Standard curve for whent coleoptile response to ARi\
St andard curve ~·or oat coleoptile response to IAA
Standard curve for oat coleoptile response to IM,
The percentage of rooting, and callusing of H'l.YW8rd cuttings treated with IHA and woundinc
The percentage rootin~ of Hayward, Abbott and Matu~ ~ropaeated under standard conditions
Hesul ts for J e 1 (y lin ,·w,lrd cutting proµagated Ull(l1'r mi~! or polythene tent
T: l'le 4. 4. 1 . 1, Results showi nf tr..e pffect of bottom
56
61
62
63
68
71
74
4. 4. 1. 2, h<::at and IBA 0.1 Jlr-'.{W,ird stem cutting 80
[l r~ri 4 . 4 . 1 . 3
•:· ., ble 4. 4 . 2. 1 Tables showi n[ the effect of
different IBA -c-oncentration on the
rooting of Abbott, stem cuttings 81 Fi['. 4.4.2.1 Percentage rootinf; of Abbott cutting 82 Fip. 4.4.2.2 The effect of v~rJous concentration
of IAA on root regeneration of
Actinidia chinensjs 83
Table 4. 5. 1 Percentage rooting on successive
harvest date 90
Fig. 4.5.1 Seasonal fluctuation in the rooting ability of Hayward ann Abbott stem
cuttings 91
Table 4.6.2.1 Percentage bud break and days to 50% bud break of cuttings of
Abbott variety 96
Fip. 4.6.2.1 Seasonal bud activities 97
Table 4.6.3.1
Fir;. 4. 7. 1
Pig. 4. 7.2
Fii-~. 4.7.3
F Lr:. 4. 7 •• 4
'I.',qble 4.8.1 T,ibJ.c 4.8.2
}'i:"'". :1 .0 .1
Fig. 4.8.3
Fip. t,.8.4
r.r a b 1 e 4 • 8 • 3
Table 4.9.1
Table 5. 2. 1
Percentage rooLing of bud and budless Abbott cuttinr,s treated with or without IRA nt various harvests
Seasonal chanr.es in rtn acidic
99
growth promoter, similar to IAA 104 Seasonal chanrcs of a neutral
growth promoter, similar to IAN 105 Seasonal changes of an acidic
growth inhibitor, s i m iJ.ar to ABA 1 06 Seasonal changes oJ' rooting
promoters from Hayw::ud cuttings as determined by the Mung Bean
Bioassay 107
Treatment for Uruno cuttinrs Precentage rootinr of Bruno cuttlng,J treated with IAA, IBA and centrifue;ai ion singly or in combination
Histograms of IAA-like growth pro:noters from stem tissue and centrifugate of Bruno
Histograms of lAN extracted from stem tisciue and centrifueate of Bruno
Histograms of ABA extracted from stem tiosue and centrifueate of Bruno
Rooting cofactors extracted from stem tissues of Bruno
Extractable Level of endogenous growth regulators from the excised bases, c1 ttings and centrifugate Bruno after centrifugation
Percentage rooting of Abbott leafy and leafless cuttings in response to IAA
Table showing the percentage shoot regeneration and number of shoot per root cutting
114
115
117
119
121 123
125
1 31
138
Fip. •,.?.1
Fig . '5.2.2
Fir-. r .2 .3
'l'ab]e r:."3.3
FiC". ' .4 .1
'l': b 1 e ~ • '1-• 1
Tab}e
5 . 5. 4
Tabel
5 .5 . 5
Table
5 . 5. 6
Fhotos showing the shoot
regeneration cc11iaci ty of roots of different di ·11neter and length Photo showing the shoot
regenernUon cnn;icity of roots with different rlinmetcr
CJ.o se-up phot o ::;how i
nr:
loc:a tion of shoot emergence from cortex of cut end.Shoot regeneration cn.·)acity of Abbot t root cutt inps taken on 11/7/77
Shoot regeneration cnpacity of Abbott root c utti :l[S tc=tken on '31/9/77
Percentage of shoot regeneration of Abbott root cuttinpo at various harvesting dater.
J>crcentage of shoot rerrencration i::1 Abbott root t·uttings at various h2rvestir1p; datP~·
Treatmerits of vc1rio•;s hormone on Abbott root cuttinrs
•rrea tments of ill\ t> and :rncrose on Abbot t ruot cut t i nr·:_;
Effect of IBA ( 1
r,o
•1wl -1 ) treatmenton Abbott root ~uttinrs
Ef .feet of GA ( SO :npl - 1
) treatment on Abbott root cuttirws
Effect of BAP ( '/?
m,
1 - 1 ) treatmenton Abbott root cuttinRs
Effect of BAP an1 sucrose on shoot regeneration
141
142
143
144
144
146
147
148
148
149
149
149
