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FOLIAR SAP NO3-N AND K OF ‘SMOOTH CAYENNE’ PINEAPPLE AND THEIR RELATIONSHIP WITH DRY MATTER YIELD

NGUYEN THANH TRIEU

SUBMITTED TO 1HE FACULTY OF THE GRADUATE SCHOOL UNIVERSITY OF THE PHILIPPINES AT LOS BANOS,

IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE

DEGREE OF

MASTER OF SCIENCE (Horticulture)

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YIELD”, prepared and submitted by NGUYEN THANH TRIEJU, in partial fulfillment of

the requirements for the degree of Master of Science (Horticulture) is hereby accepted

of eicebay feet oe ies

Member, Member,

Guidance Committee Guidance committee

Mag 26, /941 hen %, 179)

Date signed Date signed

poMito Đôn

Guidance committee

ha Voi las 9 Date signed

Accepted as partial fulfillment of requirements for the degree of Master of science

DOMINGO bus Chair, Department of Horticulture We 1⁄4 ¡(457 Dafe signed (Horticulture)

ANN INEZ N GIRONELLA

Dean, Graduate School

University of the Philippines Los Banos

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BIOGRAPHICAL SKETCH

The author was born on November 9, 1966 in Cantho province, Vietnam He is the

second child of Mr Nguyen van Day and Mrs Tu Huynh Mai

He completed his elementary education at Baven Elementary School in 1976, secondary school at Cairang Secondary School in 1980, and high school in Cairang High School in 1983 From 1983-1987, he pursued the degree of Bachelor of Science in Agriculture at University of Cantho, Cantho province Right after graduation, he was employed as Research Assistrut at the Faculty of Agriculture, Cantho University, He became instructor in 1990 up to the present

In 1994, he was granted a scholarship by Ecumenical Scholarships Programme to pursue a Master of Science Degree, Major in Horticulture with cognate in Plant Physiology (Agronomy) at the University of the Philippines at Los Banos

He is married to Nguyen thi Phuong Anh with whom he was blessed with their first son Nguyen Trong Tri

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I gratefully acknowledge Dr Rudolf Ficker, Director of Ecumenical Scholarships Programme for awarding me a scholarship to pursue a Master of Science degree in Horticulture, and Dr Tran Thuong Tuan, Vice Rector of Cantho University for granting me

a study leave

My sincere thanks and appreciation are due to Dr Domingo E Angeles, Chairman of the Department of Horticulture and Chairman of my Guidance Committee for his guidance, judicious suggestions, encouragement, understanding and generous help in conducting and writing this research work

My sincere gratitude to the other members of my Guidance Committee , Dr Flordeliza B Javier, Department of Horticulture and Dr Joveno S Lales, Department of Agronomy, for their invaluable advice, suggestions, critically review of the manuscript and

for their constructive comments

Sincere thanks are due to my friends and colleagues in the Department of Horticulture for facilitating and helping me in field and laboratory works, and others who

in one way or another contributed to this study Most special mentioned and appreciation

thanks are due to Democrito Z Magpantay, Mylene H Constantino for giving me a hand during my academic course works and thesis experiments Profound gratitude to my countrymate in UPLB, specially to Dr Phan Quang Vinh, Dr Nguyen Bao Ve, Nguyen Bao

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My profound thanks are due to all my instructors who have taught and shared their valuable knowledge to enlighten my way

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NGUYEN THANH TRIEU University of the Philippines at Los Banos, Laguna, May 1997 Foliar Sap NO;-N and_K of ‘Smooth Cayenne’ Pineapple and Their Relationship with Dry Matter Yield

Adviser: Dr Domingo E Angeles

Four experiments were conducted at the Fruit Crops Orchard of the Department of

Horticulture Department, UP Los Banos The first and the second one was done to

determine the best leaf section , leaf position and time of sampling both for NO and K, and the third and fourth aimed to determine the sensitivity of sap NO;-N and K tests to changing N and K levels, and to establish critical sap NO3-N and K at vegetative stage Incremental levels of N (0, 4, 8, 12, 16 g N per plant) and K (0, 3, 6, 9, 12 g KạO per plant) were applied to create N and K supply gradients in the soil Achlorophyllous and chlorophyllous

section of leaf B, C, D, E, F were sampled at 0800; 1000, 1200, 1400, 1600 hour and

analyzed for NO3-N at 180 and 235 days after planting, and for K at 185 and 240 days after planting The achlorophyllous section of leaf D gave the highest NO3-N from 1400 to 1600 hour The chlorophyllous section of leaf D gave better results for K; sampling can be taken at anytime of the day

In the third and fourth experiment, the sap NO3-N and K were determined at 247 and 248 days after planting Results showed that NO; test strip was sensitive to the changes of N levels in the soil Sap NO; was highly positively correlated with dry tissue NO; and total N in the plant (r = 0.91**, r = 0.82**, respectively) Dry matter yield significantly increased with the amount of N applied Sap NO;-N associated with optimum dry matter yield was 379.3 to 418.2 ppm For K, all parameters were not significantly

different

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The NO; test strip can be used to evaluate N status in pineapple Further research should be conducted to determine the relationship between sap NO3-N and fruit yield and to determine the effect of higher N levels on dry matter yield Potassium test strip was not sensitive to K application Another study can be conducted in other places where soil K is

relatively deficient

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INTRODUCTION

REVIEW OF LITERATURE

Requirement of N and K in Pineapple

Fertilizer Recommendation and Practices Evaluation of Plant Nutrient Status Factors Affecting Nutrient Level in Plant

Age of tissue

Type of tissue Kind of plant Environment

Nutrient Analysis in Pineapple Sampling method Critical nutrients

Nutrient in Plant Sap, Their Concentration and Translocation

Development of Rapid Test

Relationship Between Tissue Nitrate and Total Nitrogen Sampling Technique for Sap Test

Application of Sap Test in Other Crops

MATERIALS AND METHODS

Time and Place of the Study

Experiment 1 Sap NO;-N in Different Leaf Sections, Leaf Positions and Time of Sampling

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Experiment 2 Sap K in Different Leaf Sections, Leaf Positions

and Time of sampling

Experiment 3 Relationship Between Foliar Sap NO3-N and Dry Matter Yield

Experiment 4 Relationship Between Foliar Sap K

and Dry Matter Yield Statistical Analyses

RESULTS AND DISCUSSIONS i

Experiment 1 Sap NO;-Ñ in Different Leaf Sections, Leaf Positions and Time of Days

Sap Nitrate-N of Achlorophyllous and Chlorophyllous Section

Interaction Effect of Leaf Position,Time of Sampling and N Applied on Sap Nitrate-N in Achlorophyllous Section

Effect of Leaf Position on Sap Nitrate-N

Effect of Time of Sampling on Sap Nitrate-N Effect of Fertilizer N Level on Sap Nitrate-N

Experiment 2 Sap K Concentration in Different Leaf Sections, Leaf Positions and Time of Sampling

Sap K in Achlorophyllous and Chlorophyllous Section

Interaction Effect of Leaf Positions, Time of Sampling and K

Applied on Sap K of Chlorophyllous Section Effect of Leaf Position on Sap K

Effect of Time of Sampling on Sap K

Effect of Fertilizer Potassium Level on Sap K Experiment 3 Relationship Between Foliar Sap NO3-N

and Dry Matter Yield

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and Total N in Achlorophllous Section of Leaf D 55 Comparison of Sap and Tissue Nitrate-N 58

Relationship of Total N, Sap Nitrate-N with Dry Tissue

Nitrate-N at Various N Levels 60

Effect of Fertilizer N Level on and Relationship of Sap

Nitrate-N with Growth Parameter 62

Effect of Fertilizer N Level on and Relationship of Sap

Nitrate-N with Dry Matter Yield 64

Critical Levels of Sap Nitrate-N 64

Experiment 4 Relationship Between Foliar Sap K

and Dry Matter Yield 69

Effect of Fertilizer Potassium Level on Sap K, and Total K 69

Effect of Fertilizer K Level on the Growth Parameter 69

Effect of Fertilizer K Level on Dry Matter Yield 69

SUMMARY AND CONCLUSION 73

LITERATURE CITED 75

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LIST OF TABLES

TABLE PAGE

1 Fertilizer programs for pineapple in Guinea 6

2 Recommended fertilizer for pineapple in the Philippines 6 3 Critical nutrient levels in D-leaf of pineapple at harvest 11 4 Sap NO;-N in achlorophyllous section in pineapple leaves as affected

by different levels of nitrogen fertilizer, 180 and 235 days after planting 47 5 Sap K concentration in chlorophyllous section of leaf B, C, D, E, F in

pineapple plant, 185 and 240 days after planting 32 6 Sap K in chlorophyllous section of pineapple leaves at different time

of sampling, 185 and 240 days after planting 52

7 Sap K in chlorophyllous section of pineapple leaves as affected by

different levels of potassium fertilizer, 185 and 240 days after planting 54 8 Sap NO3-N, dry tissue NO3-N and total N in achlorophyllous section of

leaf D of pineapple, 247 days after planting 56 9 Correlation coefficients of N applied, sap NO3-N, dry tissue NO3-N,

total N 56

10 Length of leaf D, plant height, number of leaves, and diameter of pineapple plant as influenced by N application, at 247 days after

planting 63

11 Correlation coefficients of Fertilizer N level, sap NO3-N, dry tissue

NO3-N, total N and growth parameters 63

12 Effect of Fertilizer N Level on the dry matter yield of pineapple plant

as influenced by N applied, 247 days after planting 65 13 Correlation coefficients of Fertilizer N level, sap NO3-N, dry tissue

NO3-N, total N and dry mater yield 65

14 Sap K, and total K in chlorophyllous section of leaf D of pineapple

plant as effected by K application, 248 days after planting 70

15 Correlation coefficients between K applied, sap K, and total K 70

16 Length of leaf 'D, plant height, number of leaves, and diameter of plant as effected by K application, 248 days after planting 71

17 Dry matter yield of pineapple as effected by K fertilizer, 248 days after

planting 71

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FIGURE 10 11 12 13 14 15 16 17 18

The experimental area, one day after planting

Merckoquant Nitrate test kit showing the used test strip on the left

side and the unused one on the right side

Pineapple plant showing different leaf positions used in the experiment all except leaf A were sampled

Achlorophyllous and chlorophyllous sections of pineapple leaf used for sap analysis

Pineapple leaf sample is macerated using mortar and pestle

Sap is extracted and used to for testing

Result is read after one minute in the standard color scale in the test

kit

Merckoquant Kalium test kit showing the unused test strips on the left side and used ones on the right side

Test strip is dipped into the sap, and then into a reagent for one minute

Result is read using the standard color scale in the test kit

Merckoquant test strip is calibrated in the Nitracheck before it 1s dipped into the sap

Test strip is dipped in the sap extracted for one minute Sap NO; concentration is read using the Nitracheck

Sap NO;-N in achlorophyllous and chlorophyllous section of

pineapple leaves as affected by nitrogen fertilizer treatment, 180 days

after planting

Sap NO3-N in achlorophyllous and chlorophyllous section of

pineapple leaves as affected by nitrogen fertilizer treatment, 235 days after planting

Sap NO;-N achlorophyllous section of different leaf position in pineapple at different N levels, 180 days after planting

Sap NO3-N achlorophyllous section of different leaf position in pineapple at different N levels, 235 days after planting

Sap NO;-N in achlorophyllous section of pineapple leaf in relation to time of sampling, 180 days after planting

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FIGURE PAGE

19 Sap NO3-N in achlorophyllous section of pineapple leaf in relation

to time of sampling, 235 days after planting 44 20 Sap K in achlorophyllous and chlorophyllous section of pineapple

leaves in different potassium fertilizer levels, 185 days after planting 49 21 Sap K m achlorophyllous and chlorophyllous section of pineapple

leaves in different potassium fertilizer levels, 240 days after planting 50

22 Relationship of sap NO3-N concentration and N applied in pineapple,

247days after planting 57

23 Relationship of dry tissue NO3-N and sap NO3-N concentration in

achlorophyllous section of leaf in pineapple, 247days after planting 59 24 Relationship of total N and sap NO;-N concentration in

achlorophyllous section of leaf D of pineapple, 247days after planting 61

25 Relationship between total N and dry tissue NO;-N in

achlorophyllous section of leaf D of pineapple, 247 days after

planting 61

26 Relationship between plant dry weight and N application in

pineapple plant, 247 days after planting 66 27 Relationship between plant dry weight and sap NO3-N in pineapple

plant, 247 days after planting 66

28 Relationship between plant dry weight and dry tissue NO3-N in

pineapple plant, 247 days after planting 68

29 Relationship between plant dry weight and total N in pineapple plant,

247 days after planting 68

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APPENDIX A B C APPENDIX TABLE 1 10 11 12

Dry Tissue NO;- N analysig

Kjeldahl Method Used to Analyse Total N Flame photometer Method

Application of fertilizer in field experiment for development of leaf sample procedure for sap NO3-N test Application of fertilizer in field experiment for

development of leaf sample procedure for sap K test

The application of fertilizer for experiment establish

critical NO3-N level in plant sap

The application of fertilizer for experiment establish

critical K level in plant sap

Sap NO;-N concentration on achlorophyllous and chlorophyllous section of leaf B, C, D, E, F in pineapple plant, 180 and 235 days after planting

Analysis of variance for main effect and interaction among leaf position, time of sampling and fertilizer application on sap NO3-N at 180 days after planting Analysis of variance for main effect and interaction among leaf position, time of sampling and fertilizer

application on sap NO3-N at 235 days after planting

Sap NO;-N in achlorophyllous section of leaf B, C, D, E, F in pineapple, 180 and 235 days after planting

Sap NO;:-N of achlorophyllous section of leaf D in pineapple plant at different time of sampling, 180 and 235

days after planting

Test the pattern of sap NO3-N as effected by time of day at 180 days after planting

Test the pattern of sap NO3-N as effected by the time of

day at 235 days after planting

Sap K_ concentration of achlorophyllous and chlorophyllous section of leaf B, C, D, E, F in pineapple plant at 185 and 240 days after planting

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APPENDIX TABLE 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27

Analysis of variance for main effect and interaction among leaf position, time of sampling and fertilizer application on sap K at 185 days after planting

Analysis of variance for main effect and interaction among leaf position, time of sampling and fertilizer application on sap K at 240 days after planting

Analysis of variance for sap NO;-N in the

achlorophyllous section of leafD as affected by different N applied at 247 day after planting

Analysis of variance for dry tissue NO;-N in the

achlorophyllous section of leaf D as affected by different N applied at 247 day after planting

Analysis of variance for total N in the achlorophyllous section of leaf D as affected by different N applied at 247 day after planting

Analysis of variance for length of leaf D as affected by different N applied at 247 day after planting

Analysis of variance height of plant as affected by different N applied at 247 day after planting

Analysis of variance for number of leaves as affected by different N applied at 247 day after planting

Analysis of variance for diameter of plant canopy as affected by different N applied at 247 day after planting Analysis of variance for root dry weight under different N application at 247 day after planting

Analysis of variance for stem dry weight as affected by different N applied at 247 day after planting

Analysis of variance for leaves dry weight as affected by different N applied at 247 day after planting

Analysis of variance for plant dry weight as affected by different N applied at 247 day after planting

Analysis of variance for sap K concentration in chlorophyllous section of leaf D under different K

application at 248 day after planting

Analysis of variance for total K in chlorophyllous section

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K application at 248 day after planting 100 29 Analysis of variance for height of plant under different K

application at 248 day after planting 100 30 Analysis of variance for number of leaves under different

K application at 248 day after planting 101

31 Analysis of variance for diameter of plant canopy under

different K application at 248 day after planting 101

32 Analysis of variance for root dry weight under different K

application at 248 day after planting 101 33 Analysis of variance for stem dry weight under different

K application at 248 day after planting 102

34 Analysis of variance for leaves dry weight under different

K application at 248 day after planting 102 35 Analysis of variance for plant dry weight under different

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INTRODUCTION

The pineapple [Ananas comosus (L.) Merr] belongs to Bromeliaceace family It is one of the most exotic and commercialized fruit crops in the Philippines where it is grown in large areas, both for local consumption and export Its production must be increased and sustained through improved cultural management practices particularly those related to

nutrition |

Pineapple grown many years in the same land continuously removes high amount of essential nutrients from the soil If not replenished this will result in a reduction of crop yield Balanced nutrition is therefore necessary to sustain vigorous plant growth and high yield The nutritional status of a plant is influenced by various factors such as soil, fertilizer supply, relative growth rate, yield, climate, and cultural practices Nutrient deficiency or toxicity symptoms are manifested by morphological changes in the plant particularly in the leaf These visible manifestations of nutritional disorders can serve as

guide for a sound fertilization program For more precise determination of nutritional status

of the plant, regular tissue analysis should be done Result of tissue analysis will also enable the grower to correct incipient deficiencies However, tissue analysis requires precise analytical techniques It is also time consuming and laborious since it involves sending samples to the laboratory for analysis

A rapid tissue test which involves the quantitative determination of nutritional

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This study was conducted with the following objectives:

1 to establish leaf section and leaf position to sample for sap NO;-N and K analysis;

to monitor the diurnal changes in the sap NO;-N and K of the leaves;

to identify the best time of sampling;

to determine the sensitivity of sap test for NO3-N, and K to changes in the sap

nutrient concentration;

to evaluate the relationship between sap NO3-N and K in the leaves and dry matter yield as influenced by fertilization; and

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REVIEW OF LITERATURE

Pineapple is grown on sandy, sandy loam, clay loam, lateritic and peat soil Heavy clay soil is not suitable In any case the soil should be deep with good drainage and fertile Pineapple generally needs an acidic soil with a pH 5.6 to 6, but can be grown on soil with pH as high as 7.5 The optimum temperature range is from 38.8 to 50°C Although pineapple plant is fairly drought resistant, prolonged drought will seriously affect the yield

by reducing the size and number of fruits (De Geus, 1967) Requirement of N and K in Pineapple

The pineapple needs large amounts of nitrogen, and potassium The amount of N and K was removed by a yield of about 55 tons/ha (38,500 plants per ha): N: 205 kg/ha of which 43.0 was removed by fruits; K.0: 393 kg/ha of which131.0 was removed by fruits The amount removed is increased by the following amounts if one of two ratoons of each plant is removed from the field: N 24.5 kg, K.O 43 kg per ha( as cited by De Geus, 1967)

Nitrogen deficiency delays growth, stunts plant and produces yellowish green leaves The production of fruits, slips and suckers is severely affected The fruits are small and strongly colored Py et al (1957) found that nitrogen at 4 and 8 g per plant increased the height of plant, fruit weight, diameter of the fruit-bearing stem, sugar/acid ratio, pulp color scale and decreased acidity of juice, and skin color scale

Potassium requirement of pineapple is high The most typical symptom of K

deficiency is the occurrence of the small, yellow dots in the leaf tissue, either distributed

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another experiment using 6.4 and 12.4 g K20 per plant, they noted an increase in the height of plant, diameter of the fruit-bearing stem, acidity of juice, skin color scale and decreased the sugar/acid ratio, and pulp color scale

A well balanced N/K;0 ratio with at least as much KO as N should be warranted for better yield and quality For the fresh fruit market the value of N/K2O ratio is often about 1:2 For processing, N and K.O application may be as high as from 8 to 14 g N and “from 10 to 20 g K20 per plant or base on 40,000 plant per ha from 320 to 560 kg N and

from 400 to 800 kg K20 per ha

Fertilizer Recommendation and Practices

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2 gN in the form of ammonium sulphate shortly after planting, and three applications of 1g

P;O; plus 2g KạO (De Geus, 1967)

In Taiwan, Fertilizer was recommend as 16 g N, 3.2 g P20; and 16 g KạO per plant for the plant crop, and two thirds and one third of these amount for the first and the second

ratoon crop, respectively (De Geus, 1967)

In Malaya pineapple is grown on the strongly acid peaty soil in much more extensive way than in most other countries as the crop is not replanted after two or three cycles but kept for much longer period of time The main requirements are 8-10 g N and

10-12 g K;O per plant supplemented with copper Fertilizers were applied at the base of plant or as foliar spray

The Department of Agriculture in Fiji (Hall and McPaul, 1960) recommended a

mixture of 12.5 Ib urea, 15 Ib superphosphate (20% P20s) and 10 Ib K2SO, per 100 plant

for a single application The application should be repeated every four months

Py et al (1957) suggested the fertilizer programs for pineapple in Guinea (Table 1) In general, fertilizer recommendation for pineapple in Puerto Rico is 350-400 kg N, 50 -

75 kg P2Os, and 125 - 180 kg K20 applied in split applications at four month intervals In Peru, the recommended is 150-200 kg N, 60 kg P20; and 100 kg K20 per ha for pineapple

on light soils and 120-160 kg N, 40 kg P2Os, and 70 kg K20 on heavy soils Nitrogen should be applied in the form of ammonium sulphate or urea and together with potassiun,

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TIME PLANTING |, FERTILIZER QUANTITY

NUTRIENT Per plant (9) Per hecta (kg} *

For April/May N & 228

| POs 3 84 KO l1 418 Por June-Septermber N 7 266 PO; $ 132 KO 12 456 Por October/November N 8 404 Pas 4 152 E20 | 13.3 513 * Population denaty: 38,000 plante/ha

According to Angeles (1984) pineapple requires high amount of aitragencus fertilizer for vigorous growth of shoot and potash for the development of quality fruit The fertilizer recommendstion for pineapple as Table 2

Table 2 Reconunended fertilizer for pineapple in the Philippines

MONTHS APTER PLANTING NUTRIENT (2 per plant}

N K,0 i 2 3 2 - 7 2 3 107 2 - Total 8 &

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Rvaisetion of Phant lutrient States

The principal objectives of determining the nutrient status of plaxt are an follows: a} fo aid in determining the nutrient supplying power of the soil, b} to aid im determining the effect of treatment on the nutrient supply im the plant oc} fo study the relationship between the sutrient status of the plant and crop performence as an aid in predicting fertilizer requirements; and d} to help lay the foundation for approaching new probleme or for surveying unknown regions to determine where critical plant mstritional expermuentation should be conducted (Krantz et al 1948)

For plant analysic to be usefil in diagnosing the mutrients status of a plant, a port of reference must first be established This point of reference refers to the critical nutrient concentration Ulrich (1956) defined the crticml concentration as a point slightly below the concentration af maximum yield and in the transition zone between deficiency and adeauacy The eritical concentration ig mot a single value bul a narrow range of mutrient concentration above which the plant is amply supplied with mutriests and below which the

plant ic deficient (Bates, 197%}

A well defined curve describing the relationshin bebween yield sa mdtriont

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Age of tissue The effect of tissue age on nutrient concentration can be markedly affected by changes in nutrient supply (Ulrich an Hill, 1967) Any plant analysis program

must take into account the changes in nutrient content with age if it is to be usefial in

predicting the need for nutrients Thus, it is important that the physiological age of the

tissue can be the same on each plant, field or plot sampled

The change in nutrient concentration with age is probably due to increasing dry

matter and to changing proportion of certain tissue with age (Bates, 1971)

Critical concentration changes with age of tissue (Clements, 1964; Lorenz et al.,

1964) Lorenz et al (1964) sampled the petiole and rachis of the fourth leaf from the

growing tip of potato (Solanum tuberosum) petiole at 2 week intervals, and showed a rapid decline in nitrate concentration

Type of tissue The choice of plant part for sampling and analysis depends upon

the kind of plant, the purpose of the assay is to reflect the general status of the plant with

respect to the nutrient under consideration Therefore, the part selected must be of the definite physiological age and should be a uniform as possible (Thomas, 1937) Several

authors (Bould et al., 1960; Smith, 1962, and Wallace, 1961) concluded that leaves are

usually the most satisfactory part Other tissue can be used and are occasionally superior

for certain crops and nutrients ( Clements, 1964 and Emert, 1959)

Between the leaf blades and the petioles, the blade are used more frequently , but petioles have been considered superior particularly when the soluble fraction rather than

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Ulrich at al (1967) prefer the blades for Mg, Mn, Mo, K and SO.5S and the petioles for Ca, CH, and NOs-N in sugar-beets Ulrich and Hills (1967) found the petioles to be more salistkctory than blades for nitrate in sugar-beets due to the aarrower range of values obtained with the blade and due to lese sensitivity of the blades to changes in supply

The critical concentration can be directly determined only from the relation of nutrient concentration fo yield or quality and the tissue chosen should best showed this relationship

Eund of plant The critical concentration varies from species to species (Chapman,

1966) Cultivars or rootstocks and scions in the case of fruit tree and vines of a given species vary considerably in their ability to extract nutrient from soil (Epetein and Jefiric,

1964: Reather and Smith, 1954)

Kenworthy (1967) used different standard valuce of N for some apple (Avalus

spp.) cultivars then for others, the standard value representing a mean value for good plants Gowen ef al (1962) appear fo show different critical N concentration for two cultivars of cotton, Holford (1968) for N in sugarcane cultivare and Baker ef al (1966) for P im fo corn hybrids

Envionmem Bates (1971) reported a year-to-year variation in matrient

concentration and have considered weather ag a major factors mfluence nutrient component Cannel et al (1959) showed that in celery P, B and Mo concentration

decreased while Ca, Mg, and Mn increasing with increased moisture stress Fisher (1980)

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vegetalive growth when water stress wae relieved Phosphorus concentrations recover to levels similar to those im plant maintaimed in adequate moisture

Natrient Amalysis in Pineapple

needs of pineapple The commonly used leaf is D leaf (define as the youngest fully developed leaf al any time given) Critical of nutrient result of analysis leaf sampling determine the occurrig of this diagnosis fool Sideris et al (1938) recommended the basal, white, achlorophyllous portion of the D-leaf They also auggested that sample should be picked under the same climatic conditions, preferably between O800 f01000 hour on sammy days Sufficient care should be exercised when selecting the typical leaves to be picked Only one or Owo of the longest D-leaves may be sampled from any position on the plant The middle third of the white basal portion of the leaf should be used for chemical analysis

Critical nutrients Samuels et al (1958) define the optinrum leaf nutrient levels for Smooth Cayenne variety as N :1.6-1.9%6; P: 0.16-0 2086, Koi 80-2 50%

Pan (1957) defined the normal leaf mutrient values ag 1.5-2.0%N, 0.€%P and 3.5-

4.0% KE Sideris and Young (1930, 1951) established that the N content of basal parts of older leaves devoid of chlorophyll had the best correlation with the N content of the nutrient medium, while the K content of nutrient mednwn was most clearly reflected in the

green paris of the active D leaves

Godfrey-Sam-Agerey (1970) established the critical nutrient levels in D-leaf of

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11

Table 3 Critical nutrient levels in D-leaf of pineapple at harvest

LEVEL LEAF NUTRIENT IN LEAF NUTRIENT IN MIDDLE SECTION (%) BASAL SECTION (%)

N K N K

Deficient < 1.6 < 0.38 <0.35 < 0.44

Optimum 1.6-1.7 0.38 0.35-0.40 0.44

Toxic - > 0.38 > 0.40 > 0.04

Su (1961) gave the following optimum levels, applicable to Taiwan, for the basal part of the youngest leaves: 3.6% K in April (year after planting), 3.3 - 3.5% K in July, and 3.2 - 3.5% K in September In Puerto Rico, Samuels et al (1958) found a significant linear relationship between leaf potassium and yield On Coto clay, leaf potassium value near 3% was associated with optimum fruit yield, while on Lares clay at the Corozal substation the value was above 4% According to Nightingale (1942) the K content at flower differentiation should not fall below 0.38% (base on fresh material) Soil with K below 0.5 m.e per 100 g soil should definitely be considered as potash-deficient

Nutrient in Plant Sap, Their Concentration and = Translocation

Plants are known to absorb both nitrate and ammoniacal forms of nitrogen Both forms must be converted into amino acid before they are utilized by plant Ammoniacal

nitrogen does not accumulate in plant tissue, and apparently it is rapidly converted into

organic compound Nitrate nitrogen, on the other hand, does accumulate in plant and represents the chief inorganic nitrogen reserve of the tissue Thus the plant supply of

inorganic nitrogen would be related to the nitrate content of the plant sap Since the

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conducting tissue This change can be readily detected by the nitrate test The fest is mude by adding a few drops of the nitrate reagent directly on the freshly cut tissue The

presence of nitrates is indicated by development of blue color The more intense the blue

color, the greater the reserve of nitrate (Krantz et al 1948}

Organic acid and organically bound nitrogen in the form of amine acid aad smides are found in the phioem sap al the concentration between 0.2 to 0.5% Nitrate, amine acid, amides and ureides all have been implicated as principal forme of N in the xylem sap of various plants (Me Chare and Israel, 1979}

Cxthers reported that phioem exudate contains a much wider variety of organic and norganic compounds inchiding the growth substance and enzymes (Hall and Baker, 1972)

Reduced WN is incorporated into a limited number of amino acids, amides and other N solutes for transport from root to shoot (Pate, 1980} Assimilate atrogen is transported from one plant to another primarily through the phloem (Marshner, 1986} In general, mnino acid and nitrates are predominant formes of N traneport in the xylem aap (MoChure and Isracl, 1979) The ratio of amino acids and nitrate to ureides in the xylem varied depending upon the extent to which extermal N was supplied N solutes frequently comprise the major component of dry matter in xylem sap and are second only to carbohydrate in the phloem Marshner (1 986} reported that sucrose, which comprises around 90% of the solids in the phloem is the major component of phicem sap On the other hand, nitrate transport mainly occurs im the xylem Its presence, therefore in phloem exudate is not detectable tn

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Both the xylem and the khám participate in transporting N in plants, but the composition of the two saps varies widely (Pate, 1973) Gradients in hydrostatic pressure and water potential are the main driving forces for the transport of mineral elements from root to shoot which occurs predominantly in the xylem (Marshner, 1986) Solute transport in the xylem involves an exchange of adsorbed polyvalent cations and reabsorption of mineral elements followed by the release of organic compounds by surrounding cells Living cells also absorbed solute from the xylem as the xylem sap moves from root to the shoot Hence, the concentration and composition of the xylem sap changes as it moves along the plant body In the nodulated legumes, the nitrate concentration in the xylem sap decreases with increase path length (Pate et al., 1964) While long distance transport in the xylem is unidirectional, phloem transport with its living sieve tubes is bi-directional During the long distance transport mineral elements and organic solutes are transferred between the phloem and xylem by extensive exchange processes which is mediated by transfer cells (Pate and Gunning, 1972) Mobile mineral nutrients including reduced nitrogen are retranslocated from the shoots to the roots through the phloem although the roots are supplied with nutrients from the external solution (Marshner, 1986) In barley, Simpson et al (1982) found that nitrogen translocated in the xylem from the root to the shoots (100%), up to 79% was retranslocated in the phloem as reduce nitrogen back to the roots, and 21% of these reduce nitrogen was incorporated into the root tissue and the remainder translocated back in the xylem to the shoots

Development of Rapid Test

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potassium: thiocyanate The accumulation of iron in the nodes of com plant indicates K deficiency Refinement of the procechure followed through the yeara Thorton of Purdue University modified molybdate test for P and cobaltinitrate test for KE The nitrate procedure wae subsequently refined by Bray (1945) Later, Melsted (1958) developed the potash test papers using dipicrylamine

Lynd et af (1950) applied tissue fest in com, and found close correlation between

foliar analysis value and thew result of the test Their procedures require the preparation of extracts from knows amount of samples and the determination by colorimetry of the relative concentration of the nutrients

According to Ulrich, (19505 phenoldisulfome acid had been used for many years as areagent for the determination of nitrate in water, soi] extract and im plant materials In plant material, chlorides were first removed with silver sulphate since it interferes with color development The phenoldisalfonic acid was added to the oxtract The yellow color thai developed were measured with a photoelectric colorimeter

Syltie et al (1972} employed the reagent of Nelson, Kurtz and Bray known a

Bray’s nitrate powder This is a mixture of BaSO., MnSO HO, zinc, citric sotd, aulifiric

acid, and naphthylamine Any degree of red color produced on reaction with plant sap indicated the presence of nitrate

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On the other hand, McNamara et al (29715 used a dissimilatory nitrate reductase form & coli in an assay which is extremely sensitive to nitrate But this method did not become popular due to the difficulty in preparing the enzyme and the assay was Nme CORA,

Aside from using chemicals, clectrodes was also employed to determine the nitrate content im the plant issue (Paul and Carlson, 1968) According to their findings, the nitrate determmed using electrode agreed closely with nitrate determined phenoldisulfonic method

Jemison and Fox (1988) evaluated a quick test method, nitrate concentration in plant tissue and soil was measured using commercially available nitrate fest strips Quick teat regults were highly correlated with Isborstery results for both plant tissue nitrate (r = 0.87} and goi nitrate (r = 0.99} Results indicated that fest strips provide a rapid, reasonably accurate and precise method fo determine nitrate concentration in both soil and

plant fase

Several methods have also been developed for determining plants nutritional status base on its sap mutrient concentration The rapid test strip was based on colorimetric methods for the estimation of the nitrate, K level in the sap as measured againat a prepared set of coler standard Krantz et al (1948) employed this method for the field testing of corn, colton, and soybean plants using sap pressed from fresh tissue for the senmn- quantitative determination of nitrate using test papers, vial and color charts

Recently , a Merckoquant teat strip whick was initially used for testing water was

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were rolled, and within two minutes color developed The intensity of color was compared

with the standard color included in the test kit

Papastylianou, (1989).found that the use of the Merckoquant test strips offer a useful guides in deciding N fertilization of small legume cereals

Relationship Between Tissue Nitrate and Total Nitrogen

For diagnostic purposes, the most common approach has been to measure the total

tissue N by Kjeldahl procedure or to measure the nitrate in the cell sap from the fresh tissue or water soluble extract from dry tissue Total analysis reflects the summation effects with respect to nitrogen status for a period of time prior to sampling, but it fails to indicate

clearly the status at the exact time of sampling The value obtained from the total N is often

more a post-mortem than evaluation of current condition, although it is commonly used for

com and sorghum (Jones and Ecle, 1973)

On the other hand, nitrate in the tissue is indicative of current N within the plant at the time of sampling Nitrate in the tissue at a given time fails to indicate a shortage that may have occurred at an earlier stage of growth unless fertilizer was not applied and

growing condition has not appreciably changed Thus, measurement of the organic N

accumulation best reflects the cumulative effect of N supply to the plant; the inorganic form

NO3-N best reflects the current and most recent supply

Leaf nitrate value is directly correlated with nitrate content of the massive stem of the pineapple plant which may have up to half or more of its total nitrogen content in the form of nitrate The green portion of leaf contains little or no nitrate, except under

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i?

responses of plant materially However, it ie used in various metabolic reaction like nitrate reduction, oxidation of sugars, and organic nitrogen synthesia (Nightnagle, 1937)

‘The relationship between sap NOs-N and tissue NO}-N in terms of trend as affected either by fertilizer treatment or by stage of development strongly suggests that the former can be used in lieu of the latter in diagnostic analysis The relationship shows an increasing trend of sap NO:-N with that of tiseme NO;-N Scaife and Stevens (1983) oblained a close relation between tissue nitrate and sap nitrate in cabbage with R value of

9.90,

Prasad and Spiers (1984) measured nitrate N in five vegetable crops (carrot, celery, potafo, sweet corn, and tomate) at two stages of growth by fbwo methods: squeezing sap from fresh petiole or stem tissue with NO.-N determination by Merckoquant test strip; and acetic acid extraction from the dried petiole or stem with NOs-N determination by aulognalyser The relationship between hwo method was found to be highly significant with

coeflicient of determination exceeding 0.82

Nicholas (1956) compared various soluble fractions of a number of nutrients with the total concentration and in general, found that they were closely correlated except in the range of luxury consumption Ulrich (1942) stated that aitrate in sugar-beet (Seta vulgaris)

petioles reflects the N statue better than any other fraction Burham an Babiker (1968)

found nitrate in cotton {Gossypium spp.) petioles superior to the fotal N This reference for nitrate is rather conmion for tissue which accumulated aitratc

Syltic et al (1972) have correlated the result obtained in tissue tests with the result

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‘There are reports, however, that nitrate is a better indicator of N autrition than iatal N CEL Sheik ef al., 1970, Woodson and Boodley, 1983) Prasad and Spiers (1984) reported

that nitrate does not sppear inferior to the totad N Sap nitrate test was alzo found to be a

good indieator of the correlation between the current growth rate and N nutrition for sever diderent ornamental plants They found that correlation coefficients of 0.76 and 0.98 were obtamed between strip nitrate values and those from a standard isboratory procedure {acetic acid soluble NO.-N} The result indicates the accuracy of the test stip analysia and

deemed satisfactory for use in monitoring crop nitrate stafis on a routine basis Several

studies alae showed a significant linear correlation bebween sup nitrate and the reeult of isboratory analysis (Papastvlisnon, 1989}

Angeles et al (1994) used test paper sirip to determine sap NO.-N concentration in the papaya petiole and 2 standard laboratory procedure employing a nitrate electrode for dry tissue WOs-N concentration They found close correlation between sap nitrate and dry

tissue nitrate (r = 6.77) In a similar study, Tantung (1994) obtained a close correlation of r = 0.8

Sampling Technique for Sap Lest

‘The reliability of diagnostic procedures lies on the choice of samples whose

mutrient concentralion represents the actual nutrient status of plant, While the literature is

replete with recommended sarupling techniques for conventional laboratory analysis, there

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ao

of the analytical values Sample collected wrongly may result in biased or less accurate interpretation In the absence of information as to the which is the most appropriate Hesue to sample, Angeles et al (1994) used the recent mature leaves subtended with newly open flower im papaya Later, Tantung (1994) confirmed that the most recently mature leaf is the most sensifive in papaya, having the highest NO;-N compared with the youngest and eidest leafl Regardless of time of the day, in a similar vein, leaf collected at vegetative stage af 9:30 A.M was also the most sensitive

Coltman (1989) emphasized the nced for designing appropriate sampling strategy in view of high variability of petioles sap NO;s-N level from plant fo plant Besides, he reported the diurnal changes in the sap NO)-N content which was higher at 1130 hour regerdiess of days from planting

Application of Sep Test in Other Crops

Rapid tests are colorimetric methods used to determine the concentration of a

certain mutrient in plant sap base on a set of color standards Quick test has many

advantages not found in other fests I is simple, easy to do, cheap and does aot require

sophisticated equipment nor elaborate sample preparations Most importantly, it is rapid

capable of making semi-quantitaiive and precise diagnosis im less than 2 minutes Ite

application will be more important in places where government service laboratories are

lacking or non-existent Because it is simple it can be done by the farmer themeecives No technical ekill ie needed to interpret the result, if critical values are available (Angeles, 1995)

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(1983) measured the petiole sap nitrate in cabbage with the Merck test strip They found the fest fo be convenient and satisfactory Prasad and Spiers (1984) measured nitrate in five vegetable crops: carrot, celery, potato, sweet corm and tomaio using Merck test and aoanaiyser Results showed thal the relationship between fwo methods was highly significant with a coeflicient of determination exceeding 0.82

Merckoquant fest was also ueed in establishing the critical sap nitrate value for

maximam growth in kiwifruit vines (Prasad and Ravenwood, 1986) Coltman (1887) recommended that using this method in tomate requires a weekly sampling of five to {ifleen

plants to have suificient value

Merck test was also found to be a satisfactory method for determining the nitrate content of stems of cereals (Papastylisnou, 1989} and can be used in situ by flimers to predict the necessary N fertilization for maximum growth yield

'Tanhing (1994) used Merckoquant test to determine the sap NO) concentration in

the petiole of papayas, and found that petiole sap nitrate and dry tissue nitrate were highly correlated (r = 6.86) This result indicates that sap test provided a reasonable precise measure of monitoring the current nitrate concentration

Angeles (1995) evaluated the test’s sensitivity to the local init and showed that it ig capable of determining nitrate concentration in some fruit crops It can detect nitrate

concentration from the range of 50-5) ppm in jackfruit, suava, papaya, suyabano, caimito,

and atis For the fast growing crops such as cufflowers and other ornamentals and vegetables, a brighter prospect is being offered by rapid test He result can be more

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change in the supply of nutrient is more rapid and therefore the change in nutrient

concentration in plant tissue is more pronounced

Tremblay (1996) evaluated the potential of quick nitrogen test to assess N status of bean and sweet corn The test shown effective on either sap or soil extracts He concluded that quick nitrogen tests present a satisfactory approach for obtaining rapid and reliable

data on N status from either sap or soil extracts Quick nitrogen test on sweet corn tissue

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Vinee smd lace of the Study

‘The study was conducted at Proit Crops Orchard, Front Crops Division, Department of Horicultere, OP Los Banos from May 1996 to March 1997 The climate of this area belongs fo climate type I with hwo pronounced seasons, wet season from May to December and dry season from January to April From June to December 1996, mean of rainfall was 238.4 mm per month The number of rainy days was 24.1 daye per month The montha has highest amount of rainfall were July (460.5 mm with 23 ramy days} and November (406.9 mm with 25 rainy days} From January to February, mean rainfall was 26.25 om per month The temperature during the experimental period was 22.5°C to 33.3°C The relative humidity was $1-85% (National Agromet Station, UPLB} The site has Lipa clay loam soil type with good drainage, medium organic matter (2.21%), medium nitrogen {0.1656},

medium phosphorus (15.6% ppm), verv hình potasstin (2.27 ineq/100 g soil} and strongly

acidic pH (5.4) The soil was ploughed 3 times and harrowed 6 times 2 weeks before planting “Smooth Cayeme’ pineapple auckers weighing an average of 300-400 g were planted on 16 June 1996 at a distance of 1.2m 2X 0.5m XK 0.3 om in double rows (39,216 plant per ba} Before planting, suckers were treated with Alhete sokstion (0.25%), a systemic fingicide against butt and heart rot The experimental area is shown in Pig i

Three kinds of fertihzers such as urea (46% N}, sclophos (18% PO.) ammriate of potash (608 E50} were used Different levels fertilizers of these were applied depending

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Fig 2 Merckoquant Nitrate test kit showing the used test strip on the left side and the unused one on the right side

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Experiment 1 Sep NO3-N in Different Leaf Sections,

The experiment was conducted to determine the best leaf section, leaf position and time of sampling for sap nitrate using the Merckoquant Nitrate test strip (model 1-10020) iFig 2) Treatments consisting of three levels of nitrogen fertilizer were assigned fo twelve plots of 240 plants each The amount of nifrogen fertilizer application were: NO: 0, NI: 4, and N2: 8 ¢ N per plant or 0, 8.7, and 17.39 g urea, respectively Phosphate and potassium were applied al one level af 2 g P20; and 6 g K2O per plant which is equivalent to 11.11 ¢ sclophos and 10 g muriate of potash, respectively The schedule of fertilization is presented in Appendix Table 1

Leaves were sampled four weeks afler the third fertilization (180 days afler planting), and three weeks afler the fourth fertilization (235 days after planting} Leaf samples consisting of leaf B, C, D, E, F were collected from each plot af 0860, 1009,

1200, 1400, and 1600 hour The type of leaves are identified by letter designation: leaf B, old leaves near the ground, leaf C, mafure leaves, leaf D, longest filly expanded leaves; leaf E, young developing leaves, and leaf F, youngest leaves (Fig.3) The achlorophyllous section at the base of the leaf and the middle section of the chiorophylious (Pig 4) were separated from which 3-4 cm long strips were collected for sap NOs analysis

Affer sampling, the sample was immediately ground in a mortar and pestle and fransierred to a ginger prese fo extract the sap.(Fig.5 and Fig 6} The reaction zone of the Merckoquant test strip was dipped in the extracted sap (Fig 6}, Excess sap shaken off and

after one minute, read the color which developed from the reaction zone and compare this

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