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    A study of the effect of the frequency of spring cutting on the regrowth and resulting yield of lucerne (Medicago sativa L.) : a thesis presented in partial fulfillment of the requirements for the degree of Master of Agricultural Science at Massey University
    (Massey University, 1968) Tsuma, Francis
    There is abundant evidence to show that the yield, stand, chemical composition and the feeding value of lucerne may be materially affected by varying the cutting interval. Therefore, the purpose of the field experiment reported here was to determine the effect of the frequency of spring cutting on the regrowth and resulting yied of a pure stand of New Zealand certified lucerne (Chanticleer) and on weed invasion of the stand and, if possible, the reasons for this effect. It is hoped that this information will add to the existing evidence. Lucerne is cultivated for stock feed in many parts of the world, including New Zealand. In recent years, it has become increasingly important as a forage crop because of its potential for high yields of good quality feed under a wide range of climatic and soil conditions. The long tap root of the plant affords it considerable resistance to drought. Moreover, the plant is rich in phosphates, lime and protein, all of which are essential in animal production. In common with other legumes, it possesses the power to increase the nitrogen content of the soil. [FROM INTRODUCTION]
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    An investigation into some problems connected with inbreeding in lucerne (Medicago sativa) : being the results from work for a thesis for honours in Field Husbandry (M. Agr. Sc.)
    (Massey University, 1936) Driver, Charles Malcolm
    Luceme as a crop has been known for thousands of years. As far back as the history of man goes it was used in Central Asia, being the oldest plant oultivated for forage alone. It was prized by the Medians, Grecians and Romans, the Romans carrying seed with them to establish at their military bases. Where known in England in tbe 15th century, it was highly prized but was not widely known till the 17th century. It was introduced to Germany in the 16th century and America in the 19th century. Much improvement has taken place through the centuries with the result that a good variety, suited to the locality and well managed, will give a heavy yield of green herbage, when compared with the yield from pasture. This herbage is produced in the main during summer periods when it is used tor the supplementary feeding of cattle and sheep, and for the production of hay and silage.
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    A study of the effects of different frequencies of defoliation on the recovery growth of two varieties of lucerne (Medicago sativa L.) : a thesis ... for the degree of Master of Agricultural Science at Massey University.
    (Massey University, 1971) Abbott, John Munro
    In recent years there has been increasing interest in the use of lucerne in the farming systems in localities other than those which have "traditionally" grown lucerne. Much of the North Island, including the Manawatu is involved with this developing interest. To contribute to the information needed to support this, a field experiment studying the effects of different frequencies of grazing lucerne was established in 1965 at Massey University. Pure sowings of two varieties were used. These were New Zealand certified Chanticleer and New Zealand certified Wairau, with treatments ranging from continuous grazing through to hay stage defoliation. This experiment is discussed in more detail in appendix 1A. By the spring of 1969 at the commencement of the author's study, treatment differences were apparent. The author's study continued selected treatments and measured their differences of spring growth. The initial interest was in the treatment yield differences, volunteer species ingress, lucerne persistence and the associated interaction, if any, between the defoliation treatments and the spring climatic parameters. [From Introduction]
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    Studies on Stemphylium leaf spot and Leptosphaerulina pepper spot, two foliage diseases of lucerne : a thesis presented in partial fulfilment of the requirements for the degree of Masterate of Agricultural Science at Massey University
    (Massey University, 1974) Whitwell, Phillip Donald
    Over recent years the area of lucerne (Medicago sativa L.) grown in New Zealand has rapidly increased (Table 1) as farmers and commercial firms have realised the many and varied virtues of this crop in farming enterprises. TABLE 1. Areas of lucerne grown in New Zealand used for hay, silage and seed production. Year Area (acres) Authority 1953-59 144,516 N.Z. Year Book, 1963 1963-64 164,860 N.Z.Year Book,1969 1968-69 202,770 N.Z. Year Book, 1971 1970-71 300,000 Meeklah & Allen, 1971 Lynch (1967) estimated that if the present trend continued a lucerne area of at least 300,000 acres could be expected by 1977. However, this area was attained by 1970, and the increased rate of expansion can be accounted for in several ways: 1. Due to its agronomic features lucerne is primarily grown in free-draining soils subject to an unreliable summer rainfall, so ensuring a continued feed supply over this period. 2. Lucerne has a greater versatility than most crops within a farming enterprise in that it can be utilised in several ways; for example, hay and silage, grazing, seed production, and more recently for lucerne meal or pellets, and protein extraction. 3 A gross margin analysis of lucerne as a cash crop indicates a return comparable with, or better than, such other popular crops as wheat or peas (Lamb, 1969; Tocker, 1970; Anon, 1970; Anon, 1972). [From Introduction]
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    A study of herbage and seed production of lucerne (Medicago sativa L.) under different plant spacing and cutting treatments : a thesis presented in partial fulfilment of the requirements for the degree of Master of Agricultural Science at Massey University
    (Massey University, 1978) Kowithayakorn, Lumyai
    The present study on lucerne cv. "Wairau" was carried out in two separate trials, - a herbage and seed production trial and a seed development trial. The purpose of the first part was to investigate the effect of cutting and spacing on herbage production and subsequently on seed yield. In the second part, changes in seed weight, moisture content, and viability during various stages of seed development were studied. Results from the herbage study suggested that fresh and dry herbage yield were both affected more by plant spacing than by the height of cutting and the stage of plant development at the time of cutting. Close plant spacings resulted in higher yields of both dry matter per unit area and dry matter percentage than wide spacings. The study showed that lucerne plants in first year crops should be grown at a plant density higher than 44 plants per sq. metre and up to 100 plants per sq. metre to reach maximal herbage yield per unit area. Early cutting and high cutting gave a significantly lower herbage yield than when plants were cut late and cut at a low stubble height. Cutting and spacing treatments had a profound influence on seed yield per unit area. Seed production in lucerne, unlike herbage production, depended mainly on seed yield per plant rather than on the number of plants per unit area. Both cutting and spacing treatments affected seed yield per plant by altering the number of reproductive shoots, number of flowers per plant and the percentage of seed set. Plants grown at wide spacings produced more branches and flowers per plant and also exhibited a higher percentage seed set, thereby producing high numbers of seeds per plant. To obtain high seed yield in this variety of lucerne, plants should be grown at a plant density of approximately 11-25 plants per sq. metre. Cutting lucerne plants prior to allowing them to seed resulted in a weakening of the plants, severely depressed reproductive potential, delayed flowering time and subsequently affected flower production and percentage seed set. Lucerne plants should therefore not be cut at all in the year of plant establishment if high seed yields are expected. Seed development followed three distinct phases. The growth stage involved the period up to 22 days after pollination. At the end of the stage, 3% of seeds were germinable. The food reserve accumulation stage occupied the period from 23-40 days after pollination. Maximum seed dry weight was attained after 40 days at a seed moisture content of 58%. The ripening stage occupied the period from 41-75 days after pollination. At the end of this stage seed moisture content was approximately 14% and subsequently 6% of hard seed were found when seeds were tested immediately after harvest. Storage results in this trial indicated that immature seed tended to lose germination capacity more rapidly than mature seed. To ensure high seed quality in lucerne it was essential that seed be harvested after maturity.
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    The effects of temperature and irrigation on the establishment and growth of lucerne (Medicago sativa L.) on Manawatu sand country : a thesis presented in partial fulfilment of the requirements for the degree of Master of Agricultural Science in Plant Science at Massey University
    (Massey University, 1973) Brown, Noel Samuel
    This research was undertaken to study the establishment of lucerne on one of the drier soils of the Manawatu sand country. Lucerne has often been difficult to establish on these soils, yet once established grows well, possibly due to the large supply of water that is to be found in the water table a metre or so below the surface. Difficulty in establishing lucerne has often been attributed to shifting of the unconsolidated cultivated sand surface by wind, and also to the low moisture holding capacity of the soil. Lucerne stands are usually sown in the spring, but the available soil moisture may be quickly evaporated by the strong prevailing westerly wind at this time and wind erosion may occur, often burying plants with sand and leaving other young plants with their root systems exposed. Another possible reason for poor establishment of lucerne could be the high surface temperatures often experienced on the bare surface. The aim of the field study was to examine the effects of soil temperature, soil moisture and wind erosion on lucerne establishment. Subsequently, glasshouse experiments were conducted to investigate the effects of temperature variations in the soil and on the surface on the growth and survival of lucerne seedlings. [From Introduction]
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    A study of the germination and establishment of lucerne (Medicago sativa L.) oversown on an uncultivated grass sward : a thesis presented in partial fulfillment of the requirements for the degree of Master of Agricultural Science (M. Agr. Sci.) at Massey University
    (Massey University, 1977) Kusekwa, Martin Lawrence
    In this study on the oversowing of lucerne on an uncultivated sward at Massey, three trials were conducted, two in the field and one box trial under glasshouse conditions during the summer of 1975/76 and the autumn and early winter of 1976. The triars compared the effects of two lengths of ground cover, untreated, suppressed for a longer or shorter period with herbicides and burnt on the early establishment of lucerrne sown as bare or pelleted seed. It was found that small amounts of cover at sowing encouraged lucerne germination under high moisture conditions subsequent establishment and early survival was best on the burnt and chemically desiccated, plots. The cover was most useful where it was short (8 cm) and desiccated by long term paraquat or glyphosate treatments. Dense live cover was harmful to lucerne seedlings and smothered them within 5 weeks after sowing. Where recovery of the resident vegetation and weeds was rapid, as on the burnt and short term herbicide plots, lucerne establishnent and survivar was poor and surviviag plants were small and weak. Weed infestation was heavy both on burnt and herbicide treated plots, pointing to the need to introduce an early weed control programme with oversowing. Weeds were found to respond in a similar manner to lucerne to grass competition. In the autumn and early winter tria1s, slug damage was observed and this problem appeared to be of major importance under wet conditions. The effects of slugs were fourd to be most damaging during the first 4-5 weeks after sowing. Metaldehyde slug bait pellets were effective in reducing slug damage if applied at least twice during the establishment period of lucerne. The implications of these findings are reported and discussed with particular reference to the possibilities of oversowing under East African conditions
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    Studies on duration and defoliation in lucerne : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy at Massey University, New Zealand
    (Massey University, 1978) Janson, Carroll Garth
    A project was conducted to study the influence of grazing duration (GD) on lucerne Medicago sativa L. 'Wairau'. GD was defined as the period of defoliation or grazing before regrowth to the early flowering stage was again permitted. A field trial conducted for eight months from spring to autumn examined three grazing durations (GDs), 2-4, 15 and 30 days, using sheep as the grazing animal. Following this, three studies in controlled environment rooms using simulated grazing techniques allowed a more detailed study of the influence of GD and also provided an insight into the interaction of GD with climate. In both the field and the controlled environments, total herbage production for the full duration of each of the studies was always greatest under the shortest GD (0-3 days) and least under the longest GD (30 days). In the field, total herbage production was reduced by 14% under the 15 day GD system and 29% under the 30 day GD system. However in all the studies the differences in total herbage production were generated almost entirely by differences in stem yield - there were generally no treatment differences in the total production of non-stem (leaf and new shoot) material. The studies in the controlled environment rooms indicated that GD had less effect on lucerne herbage production under dry conditions than under moist conditions favouring rapid growth. Detailed shoot population studies in which large numbers of shoots were individually tagged as they arose, demonstrated the impact of shoot decapitation, the relative contributions of the different shoot types and the importance of the time of shoot appearance in relation to grazing. Differences in the immediate growth rate of the herbage following the different GDs were noted. Maximum herbage growth rates in this period followed the intermediate GDs (10-15 days) with lower growth rates after both the very short (0-3 days) and the very long (30 day) GDs. The initial regrowth inertia following the very short GD was attributed to the low number of basal shoots on this treatment at the start of the regrowth period. However the initially reduced herbage growth rates following the 30 day GDs seemed to result from an 'earlier' partitioning of assimilate to the roots in the first half of the regrowth period following this treatment. It was postulated that this partitioning effect was generated by the 'sink' effect of the depleted root system (lowest root weight, and root TNC and starch concentrations) measured at the end of a 30 day GD. The project has indicated that under active growth conditions, while GDs of 2-4 days will give maximum herbage production, GDs of 10-15 days will have little significant effect on the performance of mature sheep. Under dry conditions, or when grazing young lambs, even longer GDs of up to 30 days are unlikely to seriously affect stock production.
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    The evaluation of morphological and molecular techniques for discrimination among and verification of lucerne (Medicago sativa) cultivars : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Plant Science (Seed Technology) at Massey University, New Zealand
    (Massey University, 1996) Dehghan-Shoar, Majid; Dehghan-Shoar, Majid
    The stability and usefulness of plant morphological characters for discrimination among lucerne (Medicago sativa L.) cultivars was investigated under three sets of field and glasshouse conditions, using two New Zealand cultivars and four Iranian ecotypic cultivars representing diverse geographical adaptations. Following the guidelines of the International Union for the Protection of New Varieties of Plants (UPOV) and the requirements of the Organization for Economic Co-operation and Development (OECD) seed certification scheme for testing distinctness, homogeneity and stability of lucerne cultivars in field test plots, morphological data were recorded from four replicates of 18 spaced plants per cultivar in the field in the establishment year (1993) and the following year. Flower colour was recorded from a further 150 one-and two-year old plants of each cultivar in the field. In a glasshouse, where the minimum and maximum temperatures were set at 16°C and 22°C respectively, morphological characters were recorded from three replicates of 22 one-year old plants of each cultivar. The majority of the 12 morphological characters recommended by UPOV and OECD for discrimination and verification of lucerne cultivars were not independent of the environment. Only three characters, number of plant stems immediately above the ground, plant recovery height, and leaflet width/length ratio were stable, and thus could be used as reliable morphological descriptors for lucerne cultivars. However, none of the individual characters, or any combination of these characters, were sufficient to differentiate all of the cultivars at P<0.01, the standard required by UPOV to detect differences among lucerne cultivars. Morphologically based methods are therefore not effective for discrimination among lucerne cultivars, and there is a need to find more precise and effective techniques for assessing whether cultivars are actually different (UPOV) or whether individual seed lots of a cultivar do not differ from the cultivar standard (OECD). A digital image processing algorithm (VIPS) was used for image processing of 150 individual seeds of 17 seed lots of eight lucerne cultivars and also for image analysis of 66 leaflets from individual glasshouse grown plants of six cultivars. Of the 21 morphological characters and derived measurements recorded from individual seeds of the cultivars, 10 were useful for cultivar verification and discrimination. Among individual characters, red colour/total intensity, and blue/total intensity of seed had the highest, while actual area of seed/convex area had the lowest ability (64 % vs 18%) to discriminate among cultivars. A better cultivar discrimination (86%) was obtained on the basis of a combined value from the 10 selected characters using Canonical Discriminant Analysis. This was further improved (to 94%) using the result from the 10 individual characters plus the combination of the selected seed characters. Using image analysis of leaflets of individual plants, only 67% of the cultivars could be discriminated. Discontinuous SDS-polyacrylamide electrophoresis was used for protein analysis of both bulked and single seeds of 14 seed lots from six Iranian and two New Zealand cultivars. A series of preliminary experiments was first undertaken to determine the optimal protein loading volume for bulk, and also single seed samples of the cultivars, to enable production of clear, sharp and therefore easily evaluated bands. Of the many protein bands produced as a result of SDS-PAGE analysis of single seeds of the cultivars, the seventeen most distinctive bands were selected for analysis. Their intensities were assessed using a Vision Image Processing System (VIPS), and the data were used for discrimination among the cultivars. Electrophoresis of protein extracted from the bulked seed samples failed to differentiate the majority of the cultivars. However, 82% of the cultivars could be differentiated using the combined results from intensities of the 17 selected protein bands from single seeds of the cultivars. Since all eight cultivars could not be discriminated using SDS-PAGE of individual seeds, two other molecular techniques (Restriction Fragment Length Polymorphism (RFLP) and Random Amplified DNA Polymorphism (RAPD) were employed to determine if better discrimination among lucerne cultivars was possible using DNA techniques. DNA samples were extracted from 4-5 frozen leaves (at -70°C) from 40 individual 45 day old seedlings of six Iranian, two New Zealand, and two internationally recognised cultivars. Following purification and quantification, bulked DNA samples of the cultivars, plus those from individual seedlings were digested with four common restriction endonucleases to cleave the DNA into smaller fragments. The digested DNA fragments were separated by agarose gel electrophoresis and were then transferred to a nylon membrane. This membrane was hybridised with nine probes i.e. six lucerne cDNA probes (I013, 492, 281, 328, 473, 457), two apple probes (ADH, cDNA and ribosomal DNA (rDNA)) and one clover probe (ADH) to determine the best combination between restriction enzymes and probes to optimize the number of polymorphic bands. A non-radioactive method was used for probe labelling. Of the six lucerne probes screened with the restriction endonucleases (EcoR I, Bam HI, Hind III and Xba I) all except I013 produced extremely faint bands from the bulked DNA samples of the cultivars. Although a combination of I013 and Hind III gave the highest number of RFLP fragments in initial experiment, the result was not reproducible. No distinct RFLPs were detected using the clover ADH probe from the bulked DNA samples of the cultivars, and the apple rDNA probe detected polymorphism between only some of the cultivars. This result suggested that DNA fingerprinting of lucerne cultivars may not be feasible on the basis of the RFLPs from bulk DNA samples of the cultivars. It was therefore hypothesised that better discrimination among the cultivars might be possible on a population basis i.e. by examining the percentage of plants within each cultivar containing particular fragments. This hypothesis was tested by analysing scoring data based on the presence (1) and absence (0) of the RFLP fragments from 40 individual seedlings of each cultivar using Canonical Discriminant Analysis. RFLP analysis of individual seedlings of the cultivar using each of the rDNA and the ADH probes produced distinct but highly polymorphic RFLPs among the seedlings which was an indication of great genetic diversity within each of the cultivars. Seventy percent of cultivar pairs could be discriminated using the RFLPs detected by apple rDNA compared with 56% for the clover ADH probe at P<0.05. Some pairs of cultivars which could not be discriminated using the rDNA, were discriminated on the basis of RFLPs detected by the ADH probe. When the results from these two probes were combined, 91% of the cultivars could be discriminated. Using the PCR based RAPD technique, bulk DNA extracts from 10 lucerne cultivars plus DNA from 40 individual 45 day old seedlings of each cultivar were analyzed. Twenty-six 10-base arbitrary primers were screened using bulk DNA samples of the cultivars to select those which were able to generate clear RAPD bands. Of these, four (i.e. OPA08, OPB13 OPO19 and OPC10) produced sharp and distinctive RAPD bands. OPB19 produced the highest number of distinct RAPD fragments and all of the cultivars, even those which were closely related, could be discriminated. Although all of the cultivars could not be discriminated using the RAPD profiles generated by OPA08, OPB13 and OPC10 individually, a combination of the results from these primers provided sufficient information for discrimination among all of the 10 cultivars. The number of distinct RAPD fragments generated by a combination of the primers OPB19 and OPC10 was less than those produced by individual primers alone. To check the reproducibility of the RAPD results, replicated reactions were carried out using two primers (OPB19 and OPC10) alone and in combination with the standard reaction mixture using different batches of enzyme from the same manufacturer. This demonstrated that a high degree of reproducibility of results obtained by the RAPD technique is possible. To assess genetic variation within and among the cultivars, nine distinct RAPD bands generated by primer OPO8 were scored. The RAPD products from 40 individual seedlings of the each cultivars produced distinct but very diverse profiles, indicating genetic diversity within each individual population. Pairwise comparison of the LSMEANS of the RAPD profiles illustrated that 86% of the cultivars were significantly different. These molecular techniques and seed image analysis were also assessed for their ability to test uniformity, and detect genetic relationships among the cultivars. For seed protein and DNA analysis, uniformity was assessed using similarity of protein and DNA banding profiles from individual seeds and seedlings from each of the seed lots of the lucerne cultivars. For seed image analysis, uniformity of the cultivars was assessed on the basis of the proportion of uniform seeds in each of the seed lots. Genetic relatedness among the cultivars was detected using the squared Mahalanobis distances (D2) of the cultivars. The genetic relatedness established among the cultivars was almost the same irrespective of whether the three molecular or the seed image analysis techniques were used, despite the different nature of each analysis. The results were consistent with the genetic background, autumn-dormancy and geographical adaptation of the cultivars examined. However, these techniques did not produced the same results for testing of uniformity. Analysis of a larger sample size from a number of cultivars with known genetic background would be needed before drawing any final conclusion as to the relative merits of these techniques for estimation of relatedness and uniformity of lucerne cultivars. Comparisons between protein banding profiles and also seed morphological characters of seed lots from the same cultivar illustrated significant differences between the seed lots of some of the cultivars. This suggested that genetic shift had occurred during seed multiplication. As all of the seed lots studied had been certified under the OECD scheme, the scheme is apparently not capable of detecting this degree of genetic shift. Of all the techniques investigated, image analysis is regarded as the most suitable alterative to plot testing for discrimination among, and verification of lucerne cultivars, and for detection of genetic shift. The reasons for this are the speed of analysis, lower running costs, lack of need for chemicals and ease of operation of the equipment. However, this will be dependent on access to image analysis facilities. For genetic analysis of lucerne cultivars the RAPD technique was considered the best among the molecular techniques used.