Use of 32P in Agriculture

Chairman
DR. R. SANKAR

Presented by
S. DHARUMARAJAN 04-617-013

Isotopes

Isotopes are different forms of an atom of the same chemical element. They have identical chemical properties but a different relative atomic mass. While the number of protons is the same, the number of neutrons in the nucleus differs Radioactive nature of unstable isotopes, usually referred to as 'radioisotopes

Radioisotopes in agriculture
With the help of radioisotopes, we can easily locate the presence of a single atom and molecule and their movement. Hence, they give research workers the opportunity to follow up step by step all kinds of processes that are related to the nutrition of plant from germination to maturity. Very small quantities of labelled nutrients can be accurately measured in presence of large quantities of other nutrients. The location of materials can be identified by radio- autography. ( method of determine the distribution of radioactive material ) Tracer technique enables one in tracing those elements taken by the plants accurately and precisely. Interaction among the mineral nutrients.

Phosphorus isotopes

The chemical element phosphorus has one stable isotope (31P) and several radioisotopes (from 26P to 30P and from 32P to 38P). but only two of them (32P and 33P) are suitable for agronomic studies. Beta energy emitted by 32P is high compared to 33P The lower energy emitted by 33P presents less radiation hazard, and its longer half-life (24.4 days) allows studies to be conducted for relatively longer time periods.

Contd.,

The 32P can be easily monitored because of its high beta energy and its use is limited to P uptake studies with duration of 60 upto 90 days due to its short half-life (14.3 days).

The availability of these two isotopes also makes it possible to use double labeling techniques in root activity studies and P placement experiments

The 32P being far cheaper than obtain.

33P

and also easier and faster to

Main Characteristics of P isotopes used in plant nutrition studies

Isotopes Half-life

Radiation characteristics Type Energy

Typical applications

Exchangeable

32P

14.3 days

1.71 MeV(Emax)

P in soils P availability from P fertilizers Plant root distribution / activity Residual P fertilizer availability

33P

24.4 days

0.248 MeV(Emax)

Auto-radiography
Diffusion

in soils Double labeling with 32P

Disintegration of 32P radioisotopes

The unstable nucleus of a radioactive isotope has an excess energy To achieve stability, it undergoes random rearrangement during which energy is released in the form of particles or radiations The radioactive nuclide has excess neutrons, which is usually the case with higher elements such as 32P The stability is achieved by the conversion of neutron into proton with in the nucleus and beta particle is emitted.

n -----------------------------------------------> p+ + 32P 15---------------------------------------------> 32S 16

+ - + Energy

Basic concepts in using isotope as tracers

The first extensive use of isotopes as tracers in plant nutrition was made in the 1940s. The radioactive isotope of P (32P) was used to study the utilization of P fertilizers by various crops in a series of greenhouse experiments and field trials (Fried and Dean, 1952) A tracer was incorporated into a fertilizer material or the nutrient source of interest, and the amount of tracer taken up by the plant was measured directly. In phosphate studies, 32P carrier-free materials, i.e. contain only 32P atoms, are normally utilized as tracers. They can be also used for labeling commercial P fertilizers such as superphosphates during manufacturing or for preparing 32P labelled solutions of known concentration

contd..,

In case of natural fertilizer sources, like phosphate rocks (PR) it is not possible to directly label these materials with P isotopes, because of the changes induced in their physical and chemical characteristics during labeling.

Therefore, techniques based on reverse isotopic dilution have been widely used to investigate P availability from Phosphate rock sources to plants

Basic terminologies and definition

Radioactive tracer
A small quantity of radioisotope is used to follow a biological or chemical process.

Carrier
A quantity of stable isotopes of the element, which may be added to a radioactive isotope of that element to give a ponderable quantity to facilitate chemical operation.

Carrier free
A carrier free isotopes of an element are one in which all the atoms of the element present are radioactive.

Contd.,

Specific activity Pertaining to a radioactively sample, the amount of tracer activity (disintegrations per second) per unit amount of trace (mole)
Units of specific activity- Curie = 3.71010 disintegrating atoms per second SI unit: Becquerel 1 milli curie =37 MBq

Phosphorus in the plant derived from fertilizer (Pdff)= Specific activity of the P in the plant Specific activity of P in the fertilizer Total P in the plant derived from the fertilizer (K) Pdff total P in the plant

Percentage utilization of fertilizer of the applied fertilizer K100 Amount of applied fertilizer P

32P

isotopic techniques

Direct neutron irradiation of Phosphate rock materials Direct labeling of P fertilizer Methods based on Isotope dilution

Direct neutron irradiation of Phosphate Rock materials

It involved the incorporation of 32P by direct activation of phosphate compounds in the Phosphate rock through thermal neutron irradiation in a nuclear reactor and the use of this irradiated material as a labelled source in the evaluation of P uptake from Phosphate rock. This method has serious drawbacks such as the changes in the Phosphate rock structure, formation of non-orthophosphate 32P component and of a number of radioisotopes in the Phosphate rock matrix complicating the determinations.

This method is not adequate for evaluating Phosphate rock sources and it is not used anymore.

Direct labeling of P fertilizer

P from the fertilizer is labelled with an isotope of P (32P or 33P), it is possible to differentiate between soil- and fertilizer derived P in the plant. Thus, the amount of P taken up from the labelled P fertilizer can be measured directly.

This direct method can be used to compare several P fertilizers that can be labelled during their manufacture. This includes superphosphate, ammonium phosphates and nitro phosphates (Fried, 1954).
The efficiency of P fertilizer management practices such as timing, placement, mixing of P sources, etc; can be also measured

Methods based on Isotope dilution

Natural fertilizer materials such as phosphate rocks and organic fertilizers cannot be labelled with an isotope in a similar manner as soluble phosphates.

A method of analysis in which a known amount of tracer is added carrier free or together with w known amount of trace (carrier) to a sample containing trace of natural isotopic combination
Depending upon the system and experimental conditions used for equilibration of the isotope tracer, these measures of availability have been designated as E (Exchangeable P), L (Labile P) and A (Available P) values (Russell et al., 1954; Larsen, 1967; Fried, 1964;). General equation S1 X = ---------- X S2 X

A, L or E

Where, S1 is the specific activity of the added tracer, X is the amount of added carrier, S2 is the specific activity of tracer in the equilibrium solution.

A-value technique

The "A" value represents a quantitative measure of the availability of a soil nutrient in terms of a standard fertilizer.

The basic assumption is that when two sources of a nutrient are present in the soil, the plant will absorb from each of these sources in proportion to the respective quantities available to the plant. The amount of available (A-value) nutrient in the soil can be determined in terms of a standard, provided that the proportion of the nutrient in the plants derived from this standard is determined. This is done by using an isotopically labelled fertilizer (standard), which will give a direct measurement of the proportion of the nutrient that was derived, from the standard fertilizer Fried and Dean (1952) A= B (1-y)/y Where, A is the amount of available nutrient in the soil, B is the amount of fertilizer nutrient (standard) applied, y is the proportion of nutrient in the plant derived from fertilizer nutrient (standard).

E- Value
This method is a direct application of the isotopic dilution principle and is an attempt to measure the amount of nutrient in the soil that is in equilibrium with the same nutrient in the soil solution when 32P added to a system containing 31P, the 32P will distribute itself such that equilibrium ratio of 32P to 31P (specific activity) will be constant for all the phosphate participating in the equilibrium

Reaction
Surface 31P + solution 32P =========> surface 32P= solution 31P

At equilibrium,
Surface 31P Surface 32 P = Solution 31P Solution 32P
(Russell et al., 1954).

L- value technique

It was first suggested by Larsen (1952).

The L-value is defined as the amount of labile P (in the soil and the soil solution) that is exchangeable with 32P labelled orthophosphate ions added to the soil, as measured by a plant growing in the equilibrated soil.

The index of relative fertilizer efficiency

(LF - LO) / P application rate

Where,
LF is the L value of the fertilized treatment. LO is the L value of the treatment without P

addition.

Evaluation phosphorus use efficiency

Evaluation phosphorus use efficiency

To clearly define the optimum conditions for placement of phosphorus fertilizers To define the relative efficiency of major sources of different nutrients To achieve better understanding of the effect of time of application on the efficiencies of fertilizer. To obtain, by direct measurements, precise information on the proportion of applied fertilizer actually taken up by crops. To asses the extent of wastage that takes places if improper placement or incorrect source of fertilizer is used

P sources on drymatter production and utilization of P by rice

P- sources

Dry matter yield ( g/pot) 7.2 23.3 22.4 21.0

Total P uptake (mg/Pot)

Pdff (%)

P utilization (%)

Control DAP SSP ANP

7.5 23.4 22.9 17.5

24.9 22.5 26.8

11.8 10.4 9.4 (Dravid, 1989)

Effect of levels and sources of phosphorus on P uptake parameters in mustard

Parameters P sources SSP Dry matter uptake (g/ Pot) DAP DAP+Gypsum Mean SSP P contents (%) DAP DAP+Gypsum Mean SSP Total P uptake (mg/Pot) DAP DAP+ Gypsum Mean SSP Pdff (%) DAP DAP+ Gypsum Mean SSP Fertilizer P uptake (mg/pot) DAP DAP+ Gypsum Mean SSP P utilization (%) DAP DAP+ Gypsum Mean 2.9 0.18 5.3 0 4.8 4.4 4.7 4.6 0.26 0.24 0.29 0.28 12.3 10.4 12.2 11.6 35.8 33.6 36.3 35.3 4.4 3.5 4.4 4.1 37.5 30.0 37.7 35.0 30 5.5 4.9 5.5 5.3 0.29 0.26 0.29 0.28 15.7 13.0 16.0 14.9 38.6 35.9 38.2 37.6 6.1 4.7 6.1 5.6 26.0 19.9 26.1 24.0 P2O5 level 60 6.3 5.6 6.4 6.1 0.32 0.30 0.32 0.31 20.2 16.5 20.7 19.1 41.6 39.0 41.4 40.7 8.4 6.4 8.6 7.8 23.9 18.4 24.5 22.3 90 mean 5.5 5.0 5.5 0.29 0.26 0.29 16.1 13.3 16.3 38.7 36.2 38.6 6.3 4.9 6.3 29.1 22.7 29.4

(Sharma and Kamath, 1990)

Effect of applied P On % Pdff and % P utilization by green gram, Bengal gram, and cowpea
Dry matter yield (g/ Pot) GG 3.76 BG 3.38 C 7.15

S.No

P level (kg P2O5 ha-1) 0

Pdff (%) GG BG GG -

P utilization (%) GG BG C

30

14.6

11.2

16.9

7.4

5.0

22.2

4.40

3.84

8.72

60

25.2

17.6 24.6

8.0

4.9

20.5

5.07

4.22

9.43

90

32.4

24.1 26.9

7.4

5.1

16.7

5.03

4.42

9.81

Mean

24.1

17.6 22.8

7.6

5.0

19.8

4.56

3.97

8.78

(Thind et al., 1990)

Influence of silicates and organic amendments on dry matter yield and P uptake parameters
S.No Treatments
Dry matter yield (g/pot) 1 2 DAP DAP+ potassium silicate (100 kg/ ha) DAP+ calcium silicate (100 kg/ ha) DAP+ compost (15 tons/ha) DAP+ pressmud (15 tons/ha) SEm CD (5%) 4.50 5.80

Mustard
Total P uptake (mg/pot) 12.2 15.9 Pdff (%) P utiliza tion (%) 10.1 13.7 Dry matter yield (g/pot) 9.45 9.22

linseed
Total P uptake (mg/pot) 11.1 11.0 Pdff (%) P utiliz ation (%) 8.0 8.1

36.5 38.4

37.1 38.2

4.55

12.5

36.7

10.3

9.20

11.3

35.4

7.8

4 5

5.73 6.24 0.19 1.48

16.1 17.5 0.50 1.48

40.7 42.2 0.72 0.12

14.6 16.2 0.33 0.97

10.50 13.46 0.35 1.03

13.9 18.1 0.41 1.20

38.3 37.6 0.21 2.11

10.4 13.3 0.18 0.53

(Singh and Sharma, 1999)

Method and time of application of P use efficiency

Effect of methods of placements on Pdff and P utilization by mustard

S.No

treatments 1982-83

Pdff (%) 1983-84 17.9 24.6

P utilization (%) 1982-83 17.9 25.2 1283-1984 18.5 26.0

1 2 3 4 5

SSP broadcast SSP placement Blended SSP broadcast Blended SSP placement SSP split application CD (at 5% level)

18.3 26.5

22.4
26.0 29.5 3.6

22.3
24.1 27.8 4.0 25.9 31.7 3.1

21.4
24.6 29.9 2.7

Singh and kamath., (1990)

Effect of methods of placement on fertilizer uptake, A values and utilization by okra

S.No Treatment Fertilizer p uptake (mg/pot) Flowering 1 Placement at 5 cm depth Placement at 10 cm depth Placement at 15 cm depth Band placement one side furrow 688 Harvest 763 A values (kg P2O5/ ha) Flowering 89.8 Harvest 158 Utilization of fertilizer P (%) Flowering 11.5 Harvest 12.7 yield of fruits (Kg/pot)

3.99

763

1045

84.4

99.7

12.6

17.0

4.33

674

931

94.3

105

11.2

17.1

4.69

457 on of 688

908

99.4

119

7.6

15.3

4.51

Placements in two bands of seed furrow

708

85.7

174

11.5

11.8

4.68

(Shivananda and Iyengar., 1990)

Table 2: Utilization of P from different sources by crop 1 and 2 of rye grass as affected by method of application (Xiong et al., 1996)
Treatment CROP 1 %Pdfssp SOIL 1 Control PR uniform placement SSP uniform placement SSP fraction placement SSP-PR placement SSP-PR fraction placement SOIL 2 Control PR uniform placement SSP uniform placement SSP fraction placement uniform 90.1 87.7 91.7 %Pdfpr 2.8 %Pdfsoil 100 100 12.3 8.3 9.9 81.4 85.5 81.7 %Pdfssp CROP 2 %Pdfpr %Pdfsoil 100 100 18.6 14.5 18.3

90.4
-

5.0 95.0

6.8

80.9
-

5.4
1.1

13.7
100 98.9

86.0
90.2

14.0
9.8

81.9
84.0

18.1
16.0

SSP-PR placement
SSP-PR fraction placement

uniform

85.4
89.0

0.77
1.33

13.9
9.67

81.7
81.1

0.20
3.5

18.1
15.4

(Xiong et al., 1996)

Nutrient interaction

Effect of Nitrogen on dry matter, Pdff, and P utilization at flowering and seed yield at harvest.
N ( Kg ha-1)

S.No

Flowering stage

At harvest

Dry matter (q ha-1)

1 2 3 4 0 30 60 90 CD at 5% level 5.41 9.33 14.19 17.80 0.74

Pdff (%)

P utilization (%)
4.5 11.7 18.5 18.3 4.9

Seed yield (q ha-1)

1.54 3.12 4.24 5.64 0.10

32.6 57.1 58.3 48.3 17.2

Venkata Reddy et al., 1997

Effect of P and S levels on total P uptake, percent Pdff and fertilizer P utilization by soybean seeds.
P uptake (mg/pot)
P levels 0 25 50 mean 0

Pdff (%)
S levels 25 50 mean 0

P utilization (%)

25

50

mean

7.3

7.8

8.3

7.8

17.5

9.1

9.0

8.8

9.0

20.6

23.7

23.1

22.5

10.7

12.1

11.6

11.5

35

10.0

10.9

11.1

10.7

26.6

31.6

35.0

31.1

76

9.9

11.1

9.5

mean

8.8

9.2

9.4

23.6

27.7

29

9.2

9.2

11.0

11.4

(Khajanchi Lal et al., 1997)

P-Zn interaction on the utilization of P by wheat under varying levels of salinity.

Dry matter (g/ pot) Treatments Normal control 4.4 Saline 3.7

Total P uptake Normal 5.1 Saline 4.4

Pdff (%) Normal Saline -

P utilization (%) Normal Saline -

P P + Zn
Zn CD at 5%

11.4 15.0
9.8 0.84

9.4 13.4
10.9

19.2 23
12.8 1.54

14.4 16.1
14.6

22.0 24.9
1.62

22.1 26.4
-

10.7 14.8
1.43

8.2 10.9
-

(Dravid , 1996)

Residual effect of phosphorus

The rapid reversion of Added soluble phosphatic fertilizers is a common phenomenon in almost all soils, resulting in very low recovery, about 15- 20 per cent. Rest of the added P remains in the soil in the form of compounds which wide degree of variation in the composition, mobility and availability to the plants. Consequently large variation in the persistence of fertilizer residue and recovery ranging from 40 to 124 per cent, depending upon the number and type of subsequent crops grown. This indicates that a large portion of P remaining after the first crop is not fixed, but is indeed available to the subsequent crops. However, direct quantification of residual effect in terms of percent recovery of phosphorus is tedious and often less precise.

Effect of different levels of initially applied P on percent Pdff and fertilizer P uptake from the directly applied basal applied doses by the succeeding crops
S.No P levels ( kg P ha-1) Pdff (%) 1 2 3 4 5 0 30 60 90 mean 43.23 38.93 29.23 28.46 34.96 Green gram Fertilizer p uptake (mg P / pot) 1.89 3.85 3.78 3.16 3.17 A value (mg P / pot) 22.91 27.37 42.24 43.86 34.09 Pdff (%) 36.23 28.13 27.30 17.53 27.29 Rice Fertilizer P uptake (mg P / pot) 2.49 2.56 3.49 3.40 2.98 A value (mg P / pot) 30.71 44.58 46.47 82.09 50.96

Samaresh kundu et al., 1986

utilization of initially applied P by the component of the cropping system

P levels (kg P/ha) Wheat Fertilizer P uptake (mg P/ pot) 30 60 90 Mean 9.18 12.40 17.90 13.16 Utilization P (%) 15.30 10.33 9.44 11.85 Green gram Fertilizer P uptake (mg P/ pot) 3.55 6.69 5.49 5.24 Utiliza tion P (%) 5.91 5.57 3.03 4.83 Rice Fertilizer P uptake (mg P/ pot) 2.19 4.93 11.63 6.25 Utilization P (%) 3.65 4.10 6.45 4.73 Total P utilizati on (%)

24.86 20.00 19.42 21.42

Samaresh kundu et al., 1986

Efficiency of micro organism on P utilization

Mycorrhizal symbiosis is the most common existing symbiotic relationship in the plant kingdom. Nuclear techniques have improved the way of more precise measurements of nutritional elements absorbed by plant and where about they are transferred in the plant and soil system . 32P was used to determine the relatively more precision effect of microorganism in nutrient absorption by plant root system

Effect of P levels and vam application on Pdff, Pdfs, and phosphorus use efficiency in tomato plant
S.No treatments P uptake (mg/pot) 147 Pdff (%) 27.3 Pdfs (%) 72.7 PUE (%) 18.12

VAM control+100kg P205 SSP

VAM control+ 75 kg P205 SSP

120

26.4

73.6

19.8

VAM inoculated +100 kg P205 SSP VAM inoculated + 75 kg P205 SSP

189

33.1

66.9

28.5

140

23.1

76.9

20.2

(Dhinakaran and Savithri, 1997)

Effect of different biofertilizers on root dry weight (g plant-1), Pdff and P utilization of chickpea

S.No

Biofertilizers

root colonization P0 P60 1.5 74.8 -

Root dry weight (g plant-1) P0 0.089 0.214 0.149 P60 0.230 0.385 0.312

Pdff (%)

P utilization (%) 13.1 18.3 16.6

1 2 3

No inoculation Glomus faciculatum Pseudomonas striata

2.8 90.7 -

32.5 34.1 35

Glomus macrcarpum
mean

77.8
57.1

60.8
45.7

0.179
0.157

0.348
0.318

35.8
34.3

17.7
16.4

Mukherjee et al, (1999)

32P

in soil root environment studies

The root studies are extremely important in given soil and to device adequate cultural practices for maximizing the yield. The growth of the plants depends not only on the total quantity of nutrients present in the soil but also on the capacity of the root system to extract water and nutrients. Tracer techniques for root studies

Placement or injection of career free or labelled compounds in the soil

Plant injection technique

Placement or injection of career free or labelled compounds in the soil

32P

was introduced into the soil in angular pattern of spots at different depths from the plants.

Once the radioisotope is traced in the plant above ground, it was calculated that roots have extended to, or beyond this spot. This method is especially suited for field crops and disturbs soil minimum. In principle any radioisotope, not liable to much dispersion from the zone of placement and having nuclear characteristics of easy detection is suitable.

Plant injection technique

The amount of radioactivity proportional to the amount live roots in a certain spot. At any given stage of growth of healthy plant under a given soil, environmental conditions the ratio between root weight and shoot weight is more or less constant. If the root weight/shoot weight is proportional to specific activity of root/ specific activity of shoot, then it is possible to estimate root weight by knowing the shoot weight and the ratio of the specific activities of root and shoot.

32P

uptake is an indicator of drought tolerance

Radioisotope technique is the easiest, prcised technique to evaluate or screening the drought tolerance varieties. The aim was to relate the P uptake under moisture stress to drought tolerance. Drought tolerant cultivars had low P uptake under controlled (non stress) condition. Under moisture stress, the absorption and translocation of 32P was inhibited and the effect was more pronounced on translocation.

The severity of 32P uptake was related to moisture stress tolerance of the cultivar, lesser the inhibition more was the tolerance.
Hence, 32P uptake could be employed as an indictor of drought tolerance in developing a screening test based on radiotracer technique.

Conclusion

32P

isotopic techniques to get a better understanding of the dynamics of soil P and evaluating the agronomic effectiveness of local rock phosphate sources in cropping systems of well-defined agro-ecological zones.
32P

techniques are useful in the identification genotypes which are P deficient and Al tolerant
32P

helps to study the environmental pollution caused by phosphatic fertilizers

32P

helps to study the mechanism of mobility of P ions, i.e. rate and extent of soil-solution transfer of orthophosphate ions, in P- deficient and heavily P fertilized soils