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VI. Miscellaneous: In Vitro Propagation through Plant Tissue Culture

VI. Miscellaneous: In Vitro Propagation through Plant Tissue Culture

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PLANT CELL AND TISSUE CULTURE IN CHINA



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callus (triploid 3 n = 27) of Citrus sinensis. The diploid plantlets have been

easily induced either from bud primordia or through embryoid formation. The

triploid seedlings were regenerated through embryogenesis of endosperm calli,

which had been induced at the cell stage.

A successful system for obtaining “virus-free’’ seed potato has been worked

out by a number of institutions. Virus-free potato seed is now successfully

produced and planted in about 20 provinces. The yield has increased significantly

through the use of ‘‘virus-free’’ seed (unpublished results).

Clones of sugarcane have been obtained through tissue culture in all canegrowing provinces (unpublished results). Meristem of stem apex or lateral buds

as well as young leaves are used as initial culture materials for the induction of

callus from which young seedlings are regenerated. The yield of canes raised

through “test-tube” methods is the same as that of the control, but the tissue culture-obtained clones are more variable as some have more tillers and others have

smaller stems.

I n vitro fertilization of corn ovules was first achieved in 1977 with a simplified

medium prepared mainly from natural extracts of potato (Shao et al., 1977).

Fourteen seeds were produced in 1978, and the portion of seed set was 0.42%.

Hybrid kernels matured in about 20-22 days after pollination and germinated in

a test tube. After transferring to pots, only two plants survived. One of them had

clear purple markers on leaves and stem. The chromosome number of root cells

was 20. These results indicated that plants produced from test-tube fertilization

were intervarietal hybrids but not parthenogenetic haploids (Jiang et al., 1979).

ACKNOWLEDGMENTS

The authors wish to thank H. W.Li for his help in preparing the manuscript and R. B. Tan for her

technical assistance.



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HU HAN AND SHAO QIQUAN



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ADVANCES IN AGRONOMY. VOL. 34



HOW MUCH NITROGEN DO

LEGUMES FIX?

Thomas A. LaRue and Thomas G. Patterson

Boyce Thompson Institute for Plant Research, Ithaca, New York



I . Introduction . . . . . . . . . . . . . . . . . . . . . . . . . .

...........

A. The Importance of Obtaining Reliable Es

...........................

B. The Energy Cost of

C. Published Estimates of Nitrogen Fixation by Legume Crops . . . . . . . . . . . . . . . . . .

11. Methods of Estimating Fixation by Crops ...........................

A. Nitrogen Accumulation . . . . . . . . . . . . . . . . . . . . . . .

.........

B. Difference Methods

...............................

C. Isotopic Methods . .

...............................

D. Acetylene Reduction . . . . . . . . . . . . . . . . . . . . . . . . . .

E. Other Methods of C

111. Estimates for Major Crops . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

A. The Forages . . . . . . . . . . .

........

B. Seed Legumes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

IV. Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

A. Evaluation of Studies.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

for Future Research ..........................



........................................................



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1. INTRODUCTION

A . THEIMPORTANCEOF OBTAINING

RELIABLE

ESTIMATES



In the coming decades our increasing world population and the growing needs

for higher quality food will place severe demands on agriculture. During this

time the cost of nitrogen fertilizer will continue to rise with the price of energy.

The processes of decreasing fertility, soil erosion and desertification will force

agriculture onto land presently marginal and will boost efforts to reclaim land.

The role of legumes in agriculture is certain to increase in importance. The

pulses are sources of good-quality protein. The forages have a long-documented

history of supporting livestock on poor soil. Both these factors are attributed in

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Copyright 0 1981 by Academic Ress. Inc.

All rights of reproduction in any form reserved.

ISBN 0-12-000734-7



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THOMAS A. LARUE AND THOMAS G. PATTERSON



part to the ability of legumes, in symbiosis with rhizobia, to obtain nitrogen from

the air.

But how much nitrogen is obtained? The direction of research and the future

management of legumes in agriculture will require accurate knowledge of the

amounts fixed by crops in the field. If fixation is less than we now think, then we

will not achieve the expected returns of N. If fixation is as high or higher than

some reports say, we must search out the reason why some legume crops can

deplete soil N.

Recent work in plant physiology indicates that symbiotic fixation is not ‘‘free

fertilizer”; the plant must provide energy in the form of photosynthate. Whether

a legume crop “pays” for fixation with decreased yield is not yet determined.

Legumes require more phosphate fertilizer than cereals, and many are more

demanding of water. The cost of inoculant will not be negligible to cash-poor

farmers in developing countries. Does the N fixed compensate for these inputs?

Ultimately, plant breeders and agronomists will require accurate estimates of the

amount of fixation, and of its cost, to determine whether increasing fixation is

economically justified.

B. THEENERGYCOSTOF SYMBIOTIC

FIXATION

VERSUS

NITRATEUTILIZATION



Despite apparent differences, there is a fundamental similarity between symbiotic nitrogen fixation and the industrial production of nitrogen fertilizer.

Energy is required for both methods, for thermodynamics requires this of all

possible methods of fixation. Nitrogenase requires energy in the form of ATP

and electrons. In addition, there are energy costs associated with nodule formation and maintenance, hydrogen loss, and incorporation and transport of newly

fixed N. There must also be an energy cost for using soil nitrate, but comparisons

of the two have been difficult to examine experimentally.

Silsbury (1977, 1979) estimated the respiratory burden of subterranean clover

grown under artificial light with nitrate or NZ.He calculated the growth coefficients (the fraction of net C02uptake in light associated with the synthesis of new

dry matter) and found them constant over a 50-day period. For nodulated plants

they were significantly higher (0.189) than for plants using nitrate (0.137).

Nodulated plants used 810 mg C02 for the synthesis of 1 g dry weight, while

nonnodulated plants used only 510 mg C02.

Mahon (1977, 1978) calculated the energy cost of fixation after measuring root

respiration and estimating N fixation by acetylene reduction. He compared the

respiration of plants grown on N, with similar plants treated a few days previously with ammonium nitrate. It was assumed that the respiration due to



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